Compositions for modulating Tau expression

ABSTRACT

Disclosed herein are antisense compounds and methods for decreasing Tau mRNA and protein expression. Such methods, compounds, and compositions are useful to treat, prevent, or ameliorate Tau-associated diseases, disorders, and conditions.

SEQUENCE LISTING

The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled BIOL0227USASEQ_ST25. TXT, created Jan. 19, 2016, which is 916 Kb in size. The information in the electronic format of the sequence listing is incorporated herein by reference in its entirety.

FIELD

Provided are compositions and methods for reducing expression of Tau mRNA and protein in an animal. Such methods are useful to treat, prevent, or ameliorate neurodegenerative diseases, including Tauopathies, Alzheimer's Disease, Fronto-temporal Dementia (FTD), FTDP-17, Progressive Supranuclear Palsy (PSP), Chronic Traumatic Encephalopathy (CTE), Corticobasal Ganglionic Degeneration (CBD), Epilepsy, and Dravet's Syndrome by inhibiting expression of Tau in an animal.

BACKGROUND

The primary function of Tau is to bind to and stabilize microtubules, which are important structural components of the cytoskeleton involved in mitosis, cytokinesis, and vesicular transport. Tau is found in multiple tissues, but is particularly abundant in axons of neurons. In humans, there are six isoforms of Tau that are generated by alternative splicing of exons 2, 3, and 10. Splicing of exons 2 and 3 at the N-terminus of the protein leads to inclusion of zero, one, or two 29 amino acid acidic domains and is termed 0N, 1N, or 2N Tau respectively. The influence of these domains on Tau function is not fully clear, though may play a role in interactions with the plasma membrane. Inclusion of exon 10 at the C-terminus leads to inclusion of the microtubule binding domain encoded by exon 10. Since there are 3 microtubule binding domains elsewhere in Tau, this Tau isoform (with exon 10 included) is termed 4R Tau, where ‘R’ refers to the number of repeats of microtubule binding domains. Tau without exon 10 is termed 3R Tau. Since more microtubule binding domains (4R compared with 3R) increases the binding to microtubules, 4R Tau presumably significantly increases microtubule binding and assembly. The ratio of 3R/4R Tau is developmentally regulated, with fetal tissues expressing exclusively 3R Tau and adult human tissues expressing approximately equal levels of 3R/4R Tau. Deviations from the normal ratio of 3R/4R Tau are characteristic of neurodegenerative FTD Tauopathies. It is not known how changing the 3R/4R Tau ratio at a later stage in the adult animal will affect Tau pathogenesis.

Serine-threonine directed phosphorylation regulates the microtubule binding ability of Tau. Hyperphosphorylation promotes detachment of Tau from microtubules. Other post translational modifications of Tau have been described; however the significance of these is unclear. Phosphorylation of Tau is also developmentally regulated with higher phosphorylation in fetal tissues and much lower phosphorylation in the adult. One characteristic of neurodegenerative disorders is aberrantly increased Tau phosphorylation.

The microtubule network is involved in many important processes within the cell including structural integrity needed for maintaining morphology of cells and operating transport machinery. Since binding of Tau to microtubules stabilizes microtubules, Tau is likely to be a key mediator of some of these processes and disruption of normal Tau in neurodegenerative diseases may disrupt some of these key cellular processes.

One of the early indicators that Tau may be important in neurodegenerative syndromes was the recognition that Tau is a key component of neurofibrillary inclusions in Alzheimer's disease. In fact, neurofibrillary inclusions are aggregates of hyperphosphorylated Tau protein. Along with amyloid beta containing plaques, neurofibrillary inclusions are a hallmark of Alzheimer's disease and correlate significantly with cognitive impairment. 95% of Tau accumulations in AD are found in neuronal processes and is termed neuritic dystrophy. The process(es) whereby this microtubule associated protein becomes disengaged from microtubules and forms accumulations of proteins and how this relates to neuronal toxicity is not well understood.

Neuronal Tau inclusions are a pathological characteristic of not only Alzheimer's disease, but also a subset of Frontotemporal dementia (FTD), PSP, and CBD. The link between Tau and neurodegeneration was solidified by the discovery that mutations in the Tau gene cause a subset of FTD. These genetic data have also highlighted the importance of the 3R:4R ratio of Tau. Many of the Tau mutations that cause FTD lead to a change in Tau splicing which leads to preferential inclusion of exon 10, and thus to increased 4R Tau. The overall Tau levels are normal. Whether the Tau isoform change or the amino acid change or both cause neurodegeneration remains unknown. Recent data suggest that PSP may also be associated with an increased 4R:3R Tau ratio.

To help understand the influence of Tau ratios on neurodegeneration, a mouse model based on one of the splicing Tau mutations (N279K) has been generated using a minigene that includes the Tau promoter and the flanking intronic sequences of exon 10. As in humans, these mice demonstrate increased levels of 4R Tau compared with transgenics expressing WT Tau and develop behavioral and motor abnormalities as well as accumulations of aggregated Tau in the brain and spinal cord.

The protein “Tau” has been associated with multiple diseases of the brain including Alzheimer's disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal ganglionic degeneration, dementia pugilistica, parkinsonism linked to chromosome, Lytico-Bodig disease, tangle-predominant dementia, ganglioglioma, gangliocytoma, meningioangiomatosis, subacute sclerosing panencephalitis, lead encephalopathy, tuberous sclerosis, Hallervorden-Spatz disease, Pick's disease, argyrophilic grain disease, corticobasal degeneration or frontotemporal lobar degeneration and others. Tau-associated disorders such as AD are the most common cause of dementia in the elderly. AD affects an estimated 15 million people worldwide and 40% of the population above 85 years of age. AD is characterized by two pathological hallmarks: Tau neurofibrillary inclusions (NFT) and amyloid-β (Aβ) plaques.

There is currently a lack of acceptable options for treating such neurodegenerative diseases. It is therefore an object herein to provide methods for the treatment of such diseases.

SUMMARY

Provided herein are methods, compounds, and compositions for modulating expression of Tau mRNA and protein. In certain embodiments, compounds useful for modulating expression of Tau mRNA and protein are antisense compounds. In certain embodiments, the antisense compounds are antisense oligonucleotides.

In certain embodiments, modulation can occur in a cell or tissue. In certain embodiments, the cell or tissue is in an animal. In certain embodiments, the animal is a human. In certain embodiments, Tau mRNA levels are reduced. In certain embodiments, Tau protein levels are reduced. Such reduction can occur in a time-dependent manner or in a dose-dependent manner.

Also provided are methods, compounds, and compositions useful for preventing, treating, and ameliorating diseases, disorders, and conditions. In certain embodiments, such Tau related diseases, disorders, and conditions are neurodegenerative diseases. In certain embodiments, such neurodegenerative diseases, disorders, and conditions include Tauopathies, Alzheimer's Disease, Fronto-temporal Dementia (FTD), FTDP-17, Progressive Supranuclear Palsy (PSP), Chronic Traumatic Encephalopathy (CTE), Corticobasal Ganglionic Degeneration (CBD), Epilepsy, and Dravet's Syndrome

Such diseases, disorders, and conditions can have one or more risk factors, causes, or outcomes in common Certain risk factors and causes for development of neurodegenerative disorder include growing older, having a personal or family history, or genetic predisposition. Certain symptoms and outcomes associated with development of a neurodegenerative disorder include but are not limited to: presence of hyperphosphorylated Tau, presence of neurofibrillary inclusions, reduction of neurological function, reduced memory, reduced motor function, reduced motor coordination, and confusion.

In certain embodiments, methods of treatment include administering a Tau antisense compound to an individual in need thereof. In certain embodiments, methods of treatment include administering a Tau antisense oligonucleotide to an individual in need thereof.

The present disclosure provides the following non-limiting numbered embodiments:

Embodiment 1: A compound, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any of the nucleobase sequences of SEQ ID NOs: 20-2443 and SEQ ID NOs: 2478-2483.

Embodiment 2: A compound, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any of the nucleobase sequences of SEQ ID NOs: 2444-2477 and SEQ ID NOs: 2484-2565.

Embodiment 3: A compound, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any of the nucleobase sequences of SEQ ID NOs: 20-2565.

Embodiment 4: A compound, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and comprising a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases complementary to an equal length portion of nucleobases 135783-135980 of SEQ ID NO: 1.

Embodiment 5: A compound, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and comprising a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases complementary to an equal length portion of nucleobases 135853-135872 of SEQ ID NO: 1.

Embodiment 6: A compound, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and comprising a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases complementary to an equal length portion of nucleobases 135783-135929 of SEQ ID NO: 1.

Embodiment 7: A compound, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and comprising a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases complementary to an equal length portion of nucleobases 135783-135914 of SEQ ID NO: 1.

Embodiment 8: The compound of embodiments 4-7, wherein the nucleobase sequence of the modified oligonucleotide is at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% complementary to SEQ ID NO: 1.

Embodiment 9: The compound of any preceding embodiment, consisting of a single-stranded modified oligonucleotide.

Embodiment 10: The compound of any preceding embodiment, wherein at least one internucleoside linkage is a modified internucleoside linkage.

Embodiment 11: The compound of embodiment 10, wherein at least one modified internucleoside linkage is a phosphorothioate internucleoside linkage.

Embodiment 12: The compound of embodiment 10, wherein each modified internucleoside linkage is a phosphorothioate internucleoside linkage.

Embodiment 13: The compound of any preceding embodiment, wherein at least one internucleoside linkage is a phosphodiester internucleoside linkage.

Embodiment 14: The compound of any preceding embodiment, wherein at least one internucleoside linkage is a phosphorothioate linkage and at least one internucleoside linkage is a phosphodiester linkage.

Embodiment 15: The compound of any preceding embodiment, wherein at least one nucleoside comprises a modified nucleobase.

Embodiment 16: The compound of embodiment 15, wherein the modified nucleobase is a 5-methylcytosine

Embodiment 17: The compound of any preceding embodiment, wherein at least one nucleoside of the modified oligonucleotide comprises a modified sugar.

Embodiment 18: The compound of embodiment 17, wherein the at least one modified sugar is a bicyclic sugar.

Embodiment 19: The compound of embodiment 18, wherein the bicyclic sugar comprises a chemical link between the 2′ and 4′ position of the sugar 4′-CH2-N(R)—O-2′ bridge wherein R is, independently, H, C1-C12 alkyl, or a protecting group.

Embodiment 20: The compound of embodiment 18, wherein the bicyclic sugar comprises a 4′-CH2-N(R)—O-2′ bridge wherein R is, independently, H, C1-C12 alkyl, or a protecting group.

Embodiment 21: The compound of embodiment 17, wherein at least one modified sugar comprises a 2′-O-methoxyethyl group.

Embodiment 22: The compound of embodiment 17, wherein the modified sugar comprises a 2′-O(CH₂)₂—OCH₃ group.

Embodiment 23: The compound of any preceding embodiment, wherein the modified oligonucleotide comprises:

a gap segment consisting of 10 linked deoxynucleosides;

a 5′ wing segment consisting of 5 linked nucleosides; and

a 3′ wing segment consisting of 5 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

Embodiment 24: The compound of any preceding embodiment, wherein the modified oligonucleotide comprises:

a gap segment consisting of 9 linked deoxynucleosides;

a 5′ wing segment consisting of 5 linked nucleosides; and

a 3′ wing segment consisting of 5 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

Embodiment 25: The compound of any preceding embodiment, wherein the modified oligonucleotide comprises:

a gap segment consisting of 7 linked deoxynucleosides;

a 5′ wing segment consisting of 5 linked nucleosides; and

a 3′ wing segment consisting of 6 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

Embodiment 26: The compound of any preceding embodiment, wherein the modified oligonucleotide comprises:

a gap segment consisting of 8 linked deoxynucleosides;

a 5′ wing segment consisting of 5 linked nucleosides; and

a 3′ wing segment consisting of 5 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

Embodiment 27: The compound of any preceding embodiment, wherein the modified oligonucleotide comprises:

a gap segment consisting of 8 linked deoxynucleosides;

a 5′ wing segment consisting of 4 linked nucleosides; and

a 3′ wing segment consisting of 6 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

Embodiment 28: The compound of any preceding embodiment, wherein the modified oligonucleotide comprises:

a gap segment consisting of 8 linked deoxynucleosides;

a 5′ wing segment consisting of 6 linked nucleosides; and

a 3′ wing segment consisting of 4 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

Embodiment 29: The compound of any preceding embodiment, wherein the modified oligonucleotide consists of 20 linked nucleosides.

Embodiment 30: The compound of any preceding embodiment, wherein the modified oligonucleotide consists of 19 linked nucleosides.

Embodiment 31: The compound of any preceding embodiment, wherein the modified oligonucleotide consists of 18 linked nucleosides.

Embodiment 32: A composition comprising the compound of any preceding embodiment or salt thereof and at least one of a pharmaceutically acceptable carrier or diluent.

Embodiment 33A method comprising administering to an animal the compound or composition of any preceding embodiment.

Embodiment 34: The method of embodiment 33, wherein the animal is a human

Embodiment 35: The method of embodiment 33, wherein administering the compound prevents, treats, ameliorates, or slows progression of a tau associated disease, disorder or condition.

Embodiment 36: The method of embodiment 35, wherein the disease, disorder or condition is a Tauopathy, Alzheimer's Disease, Fronto-temporal Dementia (FTD), FTDP-17, Progressive Supranuclear Palsy (PSP), Chronic Traumatic Encephalopathy (CTE), Corticobasal Ganglionic Degeneration (CBD), Epilepsy, or Dravet's Syndrome.

Embodiment 37: Use of the compound or composition of any preceding embodiment for the manufacture of a medicament for treating a neurodegenerative disorder.

Embodiment 38: A compound consisting of ISIS 613099

Embodiment 39: A compound consisting of ISIS 613361

Embodiment 40: A compound consisting of ISIS 613370

Embodiment 41: A compound consisting of ISIS 623782

Embodiment 42: A compound consisting of ISIS 623996

Embodiment 43: A composition comprising the compound of any of embodiments 38-42, or salt thereof, and at least one of a pharmaceutically acceptable carrier or diluent.

Embodiment 44: A method comprising administering to an animal the compound or composition of any of embodiments 38-43.

Embodiment 45: The method of embodiment 44, wherein the animal is a human

Embodiment 46: The method of embodiment 44, wherein administering the compound prevents, treats, ameliorates, or slows progression of a tau associated disease, disorder or condition.

Embodiment 47: The method of embodiment 46, wherein the disease, disorder or condition is a Tauopathy, Alzheimer's Disease, Fronto-temporal Dementia (FTD), FTDP-17, Progressive Supranuclear Palsy (PSP), Chronic Traumatic Encephalopathy (CTE), Corticobasal Ganglionic Degeneration (CBD), Epilepsy, or Dravet's Syndrome.

Embodiment 48: Use of the compound or composition of any of embodiments 38-43 for the manufacture of a medicament for treating a neurodegenerative disorder.

DETAILED DESCRIPTION

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed. Herein, the use of the singular includes the plural unless specifically stated otherwise. As used herein, the use of “or” means “and/or” unless stated otherwise. Additionally, as used herein, the use of “and” means “and/or” unless stated otherwise. Furthermore, the use of the term “including” as well as other forms, such as “includes” and “included”, is not limiting. Also, terms such as “element” or “component” encompass both elements and components comprising one unit and elements and components that comprise more than one subunit, unless specifically stated otherwise.

The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents, or portions of documents, cited in this disclosure, including, but not limited to, patents, patent applications, published patent applications, articles, books, treatises, and GENBANK Accession Numbers and associated sequence information obtainable through databases such as National Center for Biotechnology Information (NCBI) and other data referred to throughout in the disclosure herein are hereby expressly incorporated by reference for the portions of the document discussed herein, as well as in their entirety.

DEFINITIONS

Unless specific definitions are provided, the nomenclature utilized in connection with, and the procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well known and commonly used in the art. Standard techniques may be used for chemical synthesis, and chemical analysis.

Unless otherwise indicated, the following terms have the following meanings:

“2′-O-methoxyethyl” (also 2′-MOE and 2′-OCH₂CH₂—OCH₃ and MOE) refers to an O-methoxyethyl modification of the 2′ position of a furanose ring. A 2′-O-methoxyethyl modified sugar is a modified sugar.

“2′-MOE nucleoside” (also 2′-O-methoxyethyl nucleoside) means a nucleoside comprising a 2′-MOE modified sugar moiety.

“2′-substituted nucleoside” means a nucleoside comprising a substituent at the 2′-position of the furanose ring other than H or OH. In certain embodiments, 2′ substituted nucleosides include nucleosides with bicyclic sugar modifications.

“5-methylcytosine” means a cytosine modified with a methyl group attached to the 5 position. A 5-methylcytosine is a modified nucleobase.

“About” means within ±7% of a value. For example, if it is stated, “the compounds affected at least about 70% inhibition of Tau”, it is implied that the Tau levels are inhibited within a range of 63% and 77%.

“Administered concomitantly” refers to the co-administration of two pharmaceutical agents in any manner in which the pharmacological effects of both are manifest in the patient at the same time. Concomitant administration does not require that both pharmaceutical agents be administered in a single pharmaceutical composition, in the same dosage form, or by the same route of administration. The effects of both pharmaceutical agents need not manifest themselves at the same time. The effects need only be overlapping for a period of time and need not be coextensive.

“Administering” means providing a pharmaceutical agent to an animal, and includes, but is not limited to administering by a medical professional and self-administering.

“Amelioration” refers to a lessening, slowing, stopping, or reversing of at least one indicator of the severity of a condition or disease. The severity of indicators may be determined by subjective or objective measures, which are known to those skilled in the art.

“Animal” refers to a human or non-human animal, including, but not limited to, mice, rats, rabbits, dogs, cats, pigs, and non-human primates, including, but not limited to, monkeys and chimpanzees.

“Antibody” refers to a molecule characterized by reacting specifically with an antigen in some way, where the antibody and the antigen are each defined in terms of the other. Antibody may refer to a complete antibody molecule or any fragment or region thereof, such as the heavy chain, the light chain, Fab region, and Fc region.

“Antisense activity” means any detectable or measurable activity attributable to the hybridization of an antisense compound to its target nucleic acid. In certain embodiments, antisense activity is a decrease in the amount or expression of a target nucleic acid or protein encoded by such target nucleic acid.

“Antisense compound” means an oligomeric compound that is capable of undergoing hybridization to a target nucleic acid through hydrogen bonding. Examples of antisense compounds include single-stranded and double-stranded compounds, such as, antisense oligonucleotides, siRNAs, shRNAs, ssRNAs, and occupancy-based compounds.

“Antisense inhibition” means reduction of target nucleic acid levels in the presence of an antisense compound complementary to a target nucleic acid compared to target nucleic acid levels or in the absence of the antisense compound.

“Antisense mechanisms” are all those mechanisms involving hybridization of a compound with a target nucleic acid, wherein the outcome or effect of the hybridization is either target degradation or target occupancy with concomitant stalling of the cellular machinery involving, for example, transcription or splicing.

“Antisense oligonucleotide” means a single-stranded oligonucleotide having a nucleobase sequence that permits hybridization to a corresponding segment of a target nucleic acid.

“Base complementarity” refers to the capacity for the precise base pairing of nucleobases of an antisense oligonucleotide with corresponding nucleobases in a target nucleic acid (i.e., hybridization), and is mediated by Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen binding between corresponding nucleobases.

“Bicyclic sugar” means a furanose ring modified by the bridging of two atoms. A bicyclic sugar is a modified sugar.

“Bicyclic nucleoside” (also BNA) means a nucleoside having a sugar moiety comprising a bridge connecting two carbon atoms of the sugar ring, thereby forming a bicyclic ring system. In certain embodiments, the bridge connects the 4′-carbon and the 2′-carbon of the sugar ring.

“Cap structure” or “terminal cap moiety” means chemical modifications, which have been incorporated at either terminus of an antisense compound.

“cEt” or “constrained ethyl” means a bicyclic nucleoside having a sugar moiety comprising a bridge connecting the 4′-carbon and the 2′-carbon, wherein the bridge has the formula: 4′-CH(CH₃)—O-2′.

“Constrained ethyl nucleoside” (also cEt nucleoside) means a nucleoside comprising a bicyclic sugar moiety comprising a 4′-CH(CH₃)—O-2′ bridge.

“Chemically distinct region” refers to a region of an antisense compound that is in some way chemically different than another region of the same antisense compound. For example, a region having 2′-O-methoxyethyl nucleosides is chemically distinct from a region having nucleosides without 2′-O-methoxyethyl modifications.

“Chimeric antisense compound” means an antisense compound that has at least two chemically distinct regions, each position having a plurality of subunits.

“Co-administration” means administration of two or more pharmaceutical agents to an individual. The two or more pharmaceutical agents may be in a single pharmaceutical composition, or may be in separate pharmaceutical compositions. Each of the two or more pharmaceutical agents may be administered through the same or different routes of administration. Co-administration encompasses parallel or sequential administration.

“Complementarity” means the capacity for pairing between nucleobases of a first nucleic acid and a second nucleic acid.

“Comprise,” “comprises,” and “comprising” will be understood to imply the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step or element or group of steps or elements.

“Contiguous nucleobases” means nucleobases immediately adjacent to each other.

“Designing” or “designed to” refer to the process of designing an oligomeric compound that specifically hybridizes with a selected nucleic acid molecule.

“Diluent” means an ingredient in a composition that lacks pharmacological activity, but is pharmaceutically necessary or desirable. For example, in drugs that are injected, the diluent may be a liquid, e.g. saline solution.

“Dose” means a specified quantity of a pharmaceutical agent provided in a single administration, or in a specified time period. In certain embodiments, a dose may be administered in one, two, or more boluses, tablets, or injections. For example, in certain embodiments where subcutaneous administration is desired, the desired dose requires a volume not easily accommodated by a single injection, therefore, two or more injections may be used to achieve the desired dose. In certain embodiments, the pharmaceutical agent is administered by infusion over an extended period of time or continuously. Doses may be stated as the amount of pharmaceutical agent per hour, day, week, or month.

“Effective amount” in the context of modulating an activity or of treating or preventing a condition means the administration of that amount of pharmaceutical agent to a subject in need of such modulation, treatment, or prophylaxis, either in a single dose or as part of a series, that is effective for modulation of that effect, or for treatment or prophylaxis or improvement of that condition. The effective amount may vary among individuals depending on the health and physical condition of the individual to be treated, the taxonomic group of the individuals to be treated, the formulation of the composition, assessment of the individual's medical condition, and other relevant factors.

“Efficacy” means the ability to produce a desired effect.

“Expression” includes all the functions by which a gene's coded information is converted into structures present and operating in a cell. Such structures include, but are not limited to the products of transcription and translation.

“Fully complementary” or “100% complementary” means each nucleobase of a first nucleic acid has a complementary nucleobase in a second nucleic acid. In certain embodiments, a first nucleic acid is an antisense compound and a target nucleic acid is a second nucleic acid.

“Gapmer” means a chimeric antisense compound in which an internal region having a plurality of nucleosides that support RNase H cleavage is positioned between external regions having one or more nucleosides, wherein the nucleosides comprising the internal region are chemically distinct from the nucleoside or nucleosides comprising the external regions. The internal region may be referred to as a “gap” and the external regions may be referred to as the “wings.”

“Gap-narrowed” means a chimeric antisense compound having a gap segment of 9 or fewer contiguous 2′-deoxyribonucleosides positioned between and immediately adjacent to 5′ and 3′ wing segments having from 1 to 6 nucleosides.

“Gap-widened” means a chimeric antisense compound having a gap segment of 12 or more contiguous 2′-deoxyribonucleosides positioned between and immediately adjacent to 5′ and 3′ wing segments having from 1 to 6 nucleosides.

“Hybridization” means the annealing of complementary nucleic acid molecules. In certain embodiments, complementary nucleic acid molecules include, but are not limited to, an antisense compound and a target nucleic acid. In certain embodiments, complementary nucleic acid molecules include, but are not limited to, an antisense oligonucleotide and a nucleic acid target.

“Identifying an animal having a Tau associated disease” means identifying an animal having been diagnosed with a Tau associated disease or predisposed to develop a Tau associated disease. Individuals predisposed to develop a Tau associated disease include those having one or more risk factors for developing a Tau associated disease, including, growing older, having a personal or family history, or genetic predisposition of one or more Tau associated diseases. Such identification may be accomplished by any method including evaluating an individual's medical history and standard clinical tests or assessments, such as genetic testing.

“Immediately adjacent” means there are no intervening elements between the immediately adjacent elements.

“Individual” means a human or non-human animal selected for treatment or therapy.

“Inhibiting Tau” means reducing the level or expression of a Tau mRNA and/or protein. In certain embodiments, Tau mRNA and/or protein levels are inhibited in the presence of an antisense compound targeting Tau, including an antisense oligonucleotide targeting Tau, as compared to expression of Tau mRNA and/or protein levels in the absence of a Tau antisense compound, such as an antisense oligonucleotide.

“Inhibiting the expression or activity” refers to a reduction or blockade of the expression or activity and does not necessarily indicate a total elimination of expression or activity.

“Internucleoside linkage” refers to the chemical bond between nucleosides.

“Linked nucleosides” means adjacent nucleosides linked together by an internucleoside linkage.

“Locked nucleic acid” or “LNA” or “LNA nucleosides” means nucleic acid monomers having a bridge connecting two carbon atoms between the 4′ and 2′position of the nucleoside sugar unit, thereby forming a bicyclic sugar. Examples of such bicyclic sugar include, but are not limited to A) α-L-Methyleneoxy (4′-CH₂—O-2′) LNA, (B) β-D-Methyleneoxy (4′-CH₂—O-2′) LNA, (C) Ethyleneoxy (4′-(CH₂)₂—O-2′) LNA, (D) Aminooxy (4′-CH₂—O—N(R)-2′) LNA and (E) Oxyamino (4′-CH₂—N(R)—O-2′) LNA, as depicted below.

As used herein, LNA compounds include, but are not limited to, compounds having at least one bridge between the 4′ and the 2′ position of the sugar wherein each of the bridges independently comprises 1 or from 2 to 4 linked groups independently selected from —[C(R₁)(R₂)]_(n)—, —C(R₁)═C(R₂)—, —C(R₁)═N—, —C(═NR₁)—, —C(═O)—, —C(═S)—, —O—, —Si(R₁)₂—, —S(═O)_(x)— and —N(R₁)—; wherein: x is 0, 1, or 2; n is 1, 2, 3, or 4; each R₁ and R₂ is, independently, H, a protecting group, hydroxyl, C₁-C₁₂ alkyl, substituted C₁-C₁₂ alkyl, C₂-C₁₂ alkenyl, substituted C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, substituted C₂-C₁₂ alkynyl, C₅-C₂₀ aryl, substituted C₅-C₂₀ aryl, a heterocycle radical, a substituted heterocycle radical, heteroaryl, substituted heteroaryl, C₅-C₇ alicyclic radical, substituted C₅-C₇ alicyclic radical, halogen, OJ₁, NJ₁J₂, SJ₁, N₃, COOJ₁, acyl (C(═O)—H), substituted acyl, CN, sulfonyl (S(═O)₂-J₁), or sulfoxyl (S(═O)-J₁); and each J₁ and J₂ is, independently, H, C₁-C₁₂ alkyl, substituted C₁-C₁₂ alkyl, C₂-C₁₂ alkenyl, substituted C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, substituted C₂-C₁₂ alkynyl, C₅-C₂₀ aryl, substituted C₅-C₂₀ aryl, acyl (C(═O)—H), substituted acyl, a heterocycle radical, a substituted heterocycle radical, C₁-C₁₂ aminoalkyl, substituted C₁-C₁₂ aminoalkyl or a protecting group.

Examples of 4′-2′ bridging groups encompassed within the definition of LNA include, but are not limited to one of formulae: —[C(R₁)(R₂)]_(n)—, —[C(R₁)(R₂)]_(n)—O—, —C(R₁R₂)—N(R₁)—O— or —C(R₁R₂)—O—N(R₁)—. Furthermore, other bridging groups encompassed with the definition of LNA are 4′-CH₂-2′, 4′-(CH₂)₂-2′, 4′-(CH₂)₃-2′, 4′-CH₂—O-2′, 4′-(CH₂)₂—O-2′, 4′-CH₂—O—N(R₁)-2′ and 4′-CH₂—N(R₁)—O-2′-bridges, wherein each R₁ and R₂ is, independently, H, a protecting group or C₁-C₁₂ alkyl.

Also included within the definition of LNA according to the invention are LNAs in which the 2′-hydroxyl group of the ribosyl sugar ring is connected to the 4′ carbon atom of the sugar ring, thereby forming a methyleneoxy (4′-CH₂—O-2′) bridge to form the bicyclic sugar moiety. The bridge can also be a methylene (—CH₂—) group connecting the 2′ oxygen atom and the 4′ carbon atom, for which the term methyleneoxy (4′-CH₂—O-2′) LNA is used. Furthermore; in the case of the bicylic sugar moiety having an ethylene bridging group in this position, the term ethyleneoxy (4′-CH₂CH₂—O-2′) LNA is used. α-L-methyleneoxy (4′-CH₂—O-2′), an isomer of methyleneoxy (4′-CH₂—O-2′) LNA is also encompassed within the definition of LNA, as used herein.

“Mismatch” or “non-complementary nucleobase” refers to the case when a nucleobase of a first nucleic acid is not capable of pairing with the corresponding nucleobase of a second or target nucleic acid.

“Modified internucleoside linkage” refers to a substitution or any change from a naturally occurring internucleoside bond (i.e., a phosphodiester internucleoside bond).

“Modified nucleobase” means any nucleobase other than adenine, cytosine, guanine, thymidine, or uracil. An “unmodified nucleobase” means the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U).

A “modified nucleoside” means a nucleoside having, independently, a modified sugar moiety and/or modified nucleobase.

“Modified nucleotide” means a nucleotide having, independently, a modified sugar moiety, modified internucleoside linkage, and/or modified nucleobase.

“Modified oligonucleotide” means an oligonucleotide comprising at least one modified internucleoside linkage, modified sugar, and/or modified nucleobase.

“Modified sugar” means substitution and/or any change from a natural sugar moiety.

“Monomer” means a single unit of an oligomer. Monomers include, but are not limited to, nucleosides and nucleotides, whether naturally occurring or modified.

“Motif” means the pattern of unmodified and modified nucleosides in an antisense compound.

“Natural sugar moiety” means a sugar moiety found in DNA (2′-H) or RNA (2′-OH).

“Naturally occurring internucleoside linkage” means a 3′ to 5′ phosphodiester linkage.

“Non-complementary nucleobase” refers to a pair of nucleobases that do not form hydrogen bonds with one another or otherwise support hybridization.

“Nucleic acid” refers to molecules composed of monomeric nucleotides. A nucleic acid includes, but is not limited to, ribonucleic acids (RNA), deoxyribonucleic acids (DNA), single-stranded nucleic acids, double-stranded nucleic acids, small interfering ribonucleic acids (siRNA), and microRNAs (miRNA).

“Nucleobase” means a heterocyclic moiety capable of pairing with a base of another nucleic acid.

“Nucleobase complementarity” refers to a nucleobase that is capable of base pairing with another nucleobase. For example, in DNA, adenine (A) is complementary to thymine (T). For example, in RNA, adenine (A) is complementary to uracil (U). In certain embodiments, complementary nucleobase refers to a nucleobase of an antisense compound that is capable of base pairing with a nucleobase of its target nucleic acid. For example, if a nucleobase at a certain position of an antisense compound is capable of hydrogen bonding with a nucleobase at a certain position of a target nucleic acid, then the position of hydrogen bonding between the oligonucleotide and the target nucleic acid is considered to be complementary at that nucleobase pair.

“Nucleobase sequence” means the order of contiguous nucleobases independent of any sugar, linkage, and/or nucleobase modification.

“Nucleoside” means a nucleobase linked to a sugar.

“Nucleoside mimetic” includes those structures used to replace the sugar or the sugar and the base and not necessarily the linkage at one or more positions of an oligomeric compound such as for example nucleoside mimetics having morpholino, cyclohexenyl, cyclohexyl, tetrahydropyranyl, bicyclo, or tricyclo sugar mimetics, e.g., non furanose sugar units. Nucleotide mimetic includes those structures used to replace the nucleoside and the linkage at one or more positions of an oligomeric compound such as for example peptide nucleic acids or morpholinos (morpholinos linked by —N(H)—C(═O)—O— or other non-phosphodiester linkage). Sugar surrogate overlaps with the slightly broader term nucleoside mimetic but is intended to indicate replacement of the sugar unit (furanose ring) only. The tetrahydropyranyl rings provided herein are illustrative of an example of a sugar surrogate wherein the furanose sugar group has been replaced with a tetrahydropyranyl ring system. “Mimetic” refers to groups that are substituted for a sugar, a nucleobase, and/or internucleoside linkage. Generally, a mimetic is used in place of the sugar or sugar-internucleoside linkage combination, and the nucleobase is maintained for hybridization to a selected target.

“Nucleotide” means a nucleoside having a phosphate group covalently linked to the sugar portion of the nucleoside.

“Off-target effect” refers to an unwanted or deleterious biological effect associated with modulation of RNA or protein expression of a gene other than the intended target nucleic acid.

“Oligomeric compound” or “oligomer” means a polymer of linked monomeric subunits which is capable of hybridizing to at least a region of a nucleic acid molecule.

“Oligonucleotide” means a polymer of linked nucleosides each of which can be modified or unmodified, independent one from another.

“Parenteral administration” means administration through injection (e.g., bolus injection) or infusion. Parenteral administration includes subcutaneous administration, intravenous administration, intramuscular administration, intraarterial administration, intraperitoneal administration, or intracranial administration, e.g., intrathecal or intracerebroventricular administration.

“Peptide” means a molecule formed by linking at least two amino acids by amide bonds. Without limitation, as used herein, peptide refers to polypeptides and proteins.

“Pharmaceutical agent” means a substance that provides a therapeutic benefit when administered to an individual. For example, in certain embodiments, an antisense oligonucleotide targeted to Tau is a pharmaceutical agent.

“Pharmaceutical composition” means a mixture of substances suitable for administering to a subject. For example, a pharmaceutical composition may comprise an antisense oligonucleotide and a sterile aqueous solution.

“Pharmaceutically acceptable derivative” encompasses pharmaceutically acceptable salts, conjugates, prodrugs or isomers of the compounds described herein.

“Pharmaceutically acceptable salts” means physiologically and pharmaceutically acceptable salts of antisense compounds, i.e., salts that retain the desired biological activity of the parent oligonucleotide and do not impart undesired toxicological effects thereto.

“Phosphorothioate linkage” means a linkage between nucleosides where the phosphodiester bond is modified by replacing one of the non-bridging oxygen atoms with a sulfur atom. A phosphorothioate linkage is a modified internucleoside linkage.

“Portion” means a defined number of contiguous (i.e., linked) nucleobases of a nucleic acid. In certain embodiments, a portion is a defined number of contiguous nucleobases of a target nucleic acid. In certain embodiments, a portion is a defined number of contiguous nucleobases of an antisense compound.

“Prevent” or “preventing” refers to delaying or forestalling the onset or development of a disease, disorder, or condition for a period of time from minutes to days, weeks to months, or indefinitely.

“Prodrug” means a therapeutic agent that is prepared in an inactive form that is converted to an active form (i.e., drug) within the body or cells thereof by the action of endogenous enzymes or other chemicals and/or conditions.

“Prophylactically effective amount” refers to an amount of a pharmaceutical agent that provides a prophylactic or preventative benefit to an animal.

“Region” is defined as a portion of the target nucleic acid having at least one identifiable structure, function, or characteristic.

“Ribonucleotide” means a nucleotide having a hydroxy at the 2′ position of the sugar portion of the nucleotide. Ribonucleotides may be modified with any of a variety of substituents.

“Salts” mean a physiologically and pharmaceutically acceptable salts of antisense compounds, i.e., salts that retain the desired biological activity of the parent oligonucleotide and do not impart undesired toxicological effects thereto.

“Segments” are defined as smaller or sub-portions of regions within a target nucleic acid.

“Shortened” or “truncated” versions of antisense oligonucleotides taught herein have one, two or more nucleosides deleted.

“Side effects” means physiological responses attributable to a treatment other than desired effects. In certain embodiments, side effects include, without limitation, injection site reactions, liver function test abnormalities, renal function abnormalities, liver toxicity, renal toxicity, central nervous system abnormalities, and myopathies.

“Single-stranded oligonucleotide” means an oligonucleotide which is not hybridized to a complementary strand.

“Sites,” as used herein, are defined as unique nucleobase positions within a target nucleic acid.

“Slows progression” means decrease in the development of the said disease.

“Specifically hybridizable” refers to an antisense compound having a sufficient degree of complementarity between an antisense oligonucleotide and a target nucleic acid to induce a desired effect, while exhibiting minimal or no effects on non-target nucleic acids under conditions in which specific binding is desired, i.e., under physiological conditions in the case of in vivo assays and therapeutic treatments.

“Stringent hybridization conditions” or “stringent conditions” refer to conditions under which an oligomeric compound will hybridize to its target sequence, but to a minimal number of other sequences.

“Subject” means a human or non-human animal selected for treatment or therapy.

“Target” refers to a protein, the modulation of which is desired.

“Target gene” refers to a gene encoding a target.

“Targeting” or “targeted” means the process of design and selection of an antisense compound that will specifically hybridize to a target nucleic acid and induce a desired effect.

“Target nucleic acid,” “target RNA,” and “target RNA transcript” and “nucleic acid target” all mean a nucleic acid capable of being targeted by antisense compounds.

“Target region” means a portion of a target nucleic acid to which one or more antisense compounds is targeted.

“Target segment” means the sequence of nucleotides of a target nucleic acid to which an antisense compound is targeted. “5′ target site” refers to the 5′-most nucleotide of a target segment. “3′ target site” refers to the 3′-most nucleotide of a target segment.

“Tau” means mammalian microtubule-associated protein tau (MAPT), including human microtubule-associated protein tau (MAPT).

“Tau associated disease” means any disease associated with any Tau nucleic acid or expression product thereof. Such diseases may include a neurodegenerative disease. Such neurodegenerative diseases may include Tauopathies, Alzheimer's Disease, Fronto-temporal Dementia (FTD), FTDP-17, Progressive Supranuclear Palsy (PSP), Chronic Traumatic Encephalopathy (CTE), Corticobasal Ganglionic Degeneration (CBD), Epilepsy, and Dravet's Syndrome.

“Tau mRNA” means any messenger RNA expression product of a DNA sequence encoding Tau.

“Tau nucleic acid” means any nucleic acid encoding Tau. For example, in certain embodiments, a Tau nucleic acid includes a DNA sequence encoding Tau, an RNA sequence transcribed from DNA encoding Tau (including genomic DNA comprising introns and exons), and an mRNA sequence encoding Tau. “Tau mRNA” means an mRNA encoding a Tau protein.

“Tau protein” means the polypeptide expression product of a Tau nucleic acid.

“Therapeutically effective amount” means an amount of a pharmaceutical agent that provides a therapeutic benefit to an individual.

“Treat” or “treating” or “treatment” refers administering a composition to effect an alteration or improvement of the disease or condition.

“Unmodified nucleobases” mean the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) and uracil (U).

“Unmodified nucleotide” means a nucleotide composed of naturally occurring nucleobases, sugar moieties, and internucleoside linkages. In certain embodiments, an unmodified nucleotide is an RNA nucleotide (i.e. β-D-ribonucleosides) or a DNA nucleotide (i.e. β-D-deoxyribonucleoside).

“Wing segment” means a plurality of nucleosides modified to impart to an oligonucleotide properties such as enhanced inhibitory activity, increased binding affinity for a target nucleic acid, or resistance to degradation by in vivo nucleases.

Certain Embodiments

Certain embodiments provide methods, compounds, and compositions for inhibiting Tau mRNA and protein expression. Certain embodiments provide methods, compounds, and composition for decreasing Tau mRNA and protein levels.

Certain embodiments provide antisense compounds targeted to a Tau nucleic acid. In certain embodiments, the Tau nucleic acid is the sequence set forth in GENBANK Accession No. GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to U.S. Pat. No. 9,381,000 (incorporated herein as SEQ ID NO: 1), GENBANK Accession No. NM_001123066.3 (incorporated herein as SEQ ID NO: 2), GENBANK Accession No. NM 016841.4, a variant mRNA sequence which skips exons 3, 4, 6, 8, 10, and 12 (incorporated herein as SEQ ID NO: 3), GENBANK Accession No. NT_010783.14 truncated from nucleotides 2624000 to U.S. Pat. No. 2,761,000 (incorporated herein as SEQ ID NO: 4), GENBANK Accession No. DR002467.1 (incorporated herein as SEQ ID NO: 5), GENBANK Accession No. NM_001203251.1 (incorporated herein as SEQ ID NO: 6), and GENBANK Accession No. NM_016835.4 (incorporated herein as SEQ ID NO: 7).

Certain embodiments provide methods for the treatment, prevention, or amelioration of diseases, disorders, and conditions associated with Tau in an individual in need thereof. Also contemplated are methods for the preparation of a medicament for the treatment, prevention, or amelioration of a disease, disorder, or condition associated with Tau. Tau associated diseases, disorders, and conditions include neurodegenerative diseases. In certain embodiments, tau associated diseases include Tauopathies, Alzheimer's Disease, Fronto-temporal Dementia (FTD), FTDP-17, Progressive Supranuclear Palsy (PSP), Chronic Traumatic Encephalopathy (CTE), Corticobasal Ganglionic Degeneration (CBD), Epilepsy, or Dravet's Syndrome.

Certain embodiments provide compounds, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any of the nucleobase sequences of SEQ ID NOs: 20-2443 and SEQ ID NOs: 2478-2483.

Certain embodiments provide compounds, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any of the nucleobase sequences of SEQ ID NOs: 2444-2477 and SEQ ID NOs: 2484-2565.

Certain embodiments provide compounds, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any of the nucleobase sequences of SEQ ID NOs: 20-2565.

Certain embodiments provide compounds, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and comprising a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases complementary to an equal length portion of nucleobases 135783-135980 of SEQ ID NO: 1.

Certain embodiments provide compounds, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and comprising a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases complementary to an equal length portion of nucleobases 135853-135872 of SEQ ID NO: 1.

Certain embodiments provide compounds, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and comprising a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases complementary to an equal length portion of nucleobases 135783-135929 of SEQ ID NO: 1.

Certain embodiments provide compounds, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and comprising a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases complementary to an equal length portion of nucleobases 135783-135914 of SEQ ID NO: 1.

In certain embodiments the nucleobase sequence of the modified oligonucleotide is at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% complementary to SEQ ID NO: 1.

In certain embodiments, the compound is a single-stranded modified oligonucleotide.

In certain embodiments, at least one internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage.

In certain embodiments, at least one modified internucleoside linkage is a phosphorothioate internucleoside linkage.

In certain embodiments, each modified internucleoside linkage is a phosphorothioate internucleoside linkage.

In certain embodiments, at least one internucleoside linkage is a phosphodiester internucleoside linkage.

In certain embodiments, at least one internucleoside linkage is a phosphorothioate linkage and at least one internucleoside linkage is a phosphodiester linkage.

In certain embodiments, at least one nucleoside comprises a modified nucleobase.

In certain embodiments, the modified nucleobase is a 5-methylcytosine.

In certain embodiments, at least one nucleoside of the modified oligonucleotide comprises a modified sugar.

In certain embodiments, at least one modified sugar is a bicyclic sugar.

In certain embodiments, the bicyclic sugar comprises a chemical link between the 2′ and 4′ position of the sugar 4′-CH2-N(R)—O-2′ bridge wherein R is, independently, H, C1-C12 alkyl, or a protecting group.

In certain embodiments, the bicyclic sugar comprises a 4′-CH2-N(R)—O-2′ bridge wherein R is, independently, H, C1-C12 alkyl, or a protecting group.

In certain embodiments, at least one modified sugar comprises a 2′-O-methoxyethyl group.

In certain embodiments, the modified sugar comprises a 2′-O(CH₂)₂—OCH₃ group.

In certain embodiments, the modified oligonucleotide comprises:

a gap segment consisting of 10 linked deoxynucleosides;

a 5′ wing segment consisting of 5 linked nucleosides; and

a 3′ wing segment consisting of 5 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

In certain embodiments, the modified oligonucleotide comprises:

a gap segment consisting of 9 linked deoxynucleosides;

a 5′ wing segment consisting of 5 linked nucleosides; and

a 3′ wing segment consisting of 5 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

In certain embodiments, the modified oligonucleotide comprises:

a gap segment consisting of 7 linked deoxynucleosides;

a 5′ wing segment consisting of 5 linked nucleosides; and

a 3′ wing segment consisting of 6 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

In certain embodiments, the modified oligonucleotide comprises:

a gap segment consisting of 8 linked deoxynucleosides;

a 5′ wing segment consisting of 5 linked nucleosides; and

a 3′ wing segment consisting of 5 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

In certain embodiments, the modified oligonucleotide comprises:

a gap segment consisting of 8 linked deoxynucleosides;

a 5′ wing segment consisting of 4 linked nucleosides; and

a 3′ wing segment consisting of 6 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

In certain embodiments, the modified oligonucleotide comprises:

a gap segment consisting of 8 linked deoxynucleosides;

a 5′ wing segment consisting of 6 linked nucleosides; and

a 3′ wing segment consisting of 4 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

In certain embodiments, the modified oligonucleotide consists of 20 linked nucleosides.

In certain embodiments, the modified oligonucleotide consists of 19 linked nucleosides.

In certain embodiments, the modified oligonucleotide consists of 18 linked nucleosides.

Certain embodiments provide compositions comprising any compound described herein or salt thereof and at least one of a pharmaceutically acceptable carrier or diluent.

Certain embodiments provide methods comprising administering to an animal any compound or composition described herein.

In certain embodiments, the animal is a human.

In certain embodiments, administering the compound prevents, treats, ameliorates, or slows progression of a tau associated disease, disorder or condition.

In certain embodiments, the disease, disorder or condition is a Tauopathy, Alzheimer's Disease, Fronto-temporal Dementia (FTD), FTDP-17, Progressive Supranuclear Palsy (PSP), Chronic Traumatic Encephalopathy (CTE), Corticobasal Ganglionic Degeneration (CBD), Epilepsy, or Dravet's Syndrome.

Certain embodiments provide use of any of the compounds or compositions of described herein for the manufacture of a medicament for treating a neurodegenerative disorder.

Certain embodiments provide compounds according to the following formula (Ia):

or a pharmaceutically acceptable salt thereof. In certain embodiments, provided are pharmaceutical compositions comprising the compound having the formula (Ia).

Certain embodiments provide compounds according to the following formula (IIa):

or a pharmaceutically acceptable salt thereof. In certain embodiments, provided are pharmaceutical compositions comprising the compound having the formula (IIa).

Certain embodiments provide compounds according to the following formula (Ma):

or a pharmaceutically acceptable salt thereof. In certain embodiments, provided are pharmaceutical compositions comprising the compound having the formula (Ma).

Certain embodiments provide compounds according to the following formula (IVa):

or a pharmaceutically acceptable salt thereof. In certain embodiments, provided are pharmaceutical compositions comprising the compound having the formula (IVa).

Certain embodiments provide compounds according to the following formula (Va):

or a pharmaceutically acceptable salt thereof. In certain embodiments, provided are pharmaceutical compositions comprising the compound having the formula (Va). Antisense Compounds

Oligomeric compounds include, but are not limited to, oligonucleotides, oligonucleosides, oligonucleotide analogs, oligonucleotide mimetics, antisense compounds, antisense oligonucleotides, and siRNAs. An oligomeric compound may be “antisense” to a target nucleic acid, meaning that is capable of undergoing hybridization to a target nucleic acid through hydrogen bonding.

In certain embodiments, an antisense compound has a nucleobase sequence that, when written in the 5′ to 3′ direction, comprises the reverse complement of the target segment of a target nucleic acid to which it is targeted. In certain such embodiments, an antisense oligonucleotide has a nucleobase sequence that, when written in the 5′ to 3′ direction, comprises the reverse complement of the target segment of a target nucleic acid to which it is targeted.

In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 12 to 30 subunits in length. In certain embodiments, an antisense compound targeted to Tau nucleic acid is 12 to 25 subunits in length. In certain embodiments, an antisense compound targeted to Tau nucleic acid is 12 to 22 subunits in length. In certain embodiments, an antisense compound targeted to Tau nucleic acid is 14 to 20 subunits in length. In certain embodiments, an antisense compound targeted to Tau nucleic acid is 15 to 25 subunits in length. In certain embodiments, an antisense compound targeted to Tau nucleic acid is 18 to 22 subunits in length. In certain embodiments, an antisense compound targeted to Tau nucleic acid is 19 to 21 subunits in length. In certain embodiments, the antisense compound is 8 to 80, 12 to 50, 13 to 30, 13 to 50, 14 to 30, 14 to 50, 15 to 30, 15 to 50, 16 to 30, 16 to 50, 17 to 30, 17 to 50, 18 to 30, 18 to 50, 19 to 30, 19 to 50, or 20 to 30 linked subunits in length.

In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 12 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 13 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 14 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 15 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 16 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 17 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 18 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 19 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 20 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 21 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 22 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 23 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 24 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 25 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 26 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 27 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 28 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 29 subunits in length. In certain embodiments, an antisense compound targeted to a Tau nucleic acid is 30 subunits in length. In certain embodiments, the antisense compound targeted to a Tau nucleic acid is 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, or 80 linked subunits in length, or a range defined by any two of the above values. In certain embodiments the antisense compound is an antisense oligonucleotide, and the linked subunits are nucleosides.

In certain embodiments antisense oligonucleotides targeted to a Tau nucleic acid may be shortened or truncated. For example, a single subunit may be deleted from the 5′ end (5′ truncation), or alternatively from the 3′ end (3′ truncation). A shortened or truncated antisense compound targeted to a Tau nucleic acid may have two subunits deleted from the 5′ end, or alternatively may have two subunits deleted from the 3′ end, of the antisense compound. Alternatively, the deleted nucleosides may be dispersed throughout the antisense compound, for example, in an antisense compound having one nucleoside deleted from the 5′ end and one nucleoside deleted from the 3′ end.

When a single additional subunit is present in a lengthened antisense compound, the additional subunit may be located at the 5′ or 3′ end of the antisense compound. When two or more additional subunits are present, the added subunits may be adjacent to each other, for example, in an antisense compound having two subunits added to the 5′ end (5′ addition), or alternatively to the 3′ end (3′ addition), of the antisense compound. Alternatively, the added subunits may be dispersed throughout the antisense compound, for example, in an antisense compound having one subunit added to the 5′ end and one subunit added to the 3′ end.

It is possible to increase or decrease the length of an antisense compound, such as an antisense oligonucleotide, and/or introduce mismatch bases without eliminating activity. For example, in Woolf et al. (Proc. Natl. Acad. Sci. USA 89:7305-7309, 1992), a series of antisense oligonucleotides 13-25 nucleobases in length were tested for their ability to induce cleavage of a target RNA in an oocyte injection model. Antisense oligonucleotides 25 nucleobases in length with 8 or 11 mismatch bases near the ends of the antisense oligonucleotides were able to direct specific cleavage of the target mRNA, albeit to a lesser extent than the antisense oligonucleotides that contained no mismatches. Similarly, target specific cleavage was achieved using 13 nucleobase antisense oligonucleotides, including those with 1 or 3 mismatches.

Gautschi et al (J. Natl. Cancer Inst. 93:463-471, March 2001) demonstrated the ability of an oligonucleotide having 100% complementarity to the bcl-2 mRNA and having 3 mismatches to the bcl-xL mRNA to reduce the expression of both bcl-2 and bcl-xL in vitro and in vivo. Furthermore, this oligonucleotide demonstrated potent anti-tumor activity in vivo.

Maher and Dolnick (Nuc. Acid. Res. 16:3341-3358, 1988) tested a series of tandem 14 nucleobase antisense oligonucleotides, and a 28 and 42 nucleobase antisense oligonucleotides comprised of the sequence of two or three of the tandem antisense oligonucleotides, respectively, for their ability to arrest translation of human DHFR in a rabbit reticulocyte assay. Each of the three 14 nucleobase antisense oligonucleotides alone was able to inhibit translation, albeit at a more modest level than the 28 or 42 nucleobase antisense oligonucleotides.

Antisense Compound Motifs

In certain embodiments, antisense compounds targeted to a Tau nucleic acid have chemically modified subunits arranged in patterns, or motifs, to confer to the antisense compounds properties such as enhanced inhibitory activity, increased binding affinity for a target nucleic acid, or resistance to degradation by in vivo nucleases.

Chimeric antisense compounds typically contain at least one region modified so as to confer increased resistance to nuclease degradation, increased cellular uptake, increased binding affinity for the target nucleic acid, and/or increased inhibitory activity. A second region of a chimeric antisense compound may optionally serve as a substrate for the cellular endonuclease RNase H, which cleaves the RNA strand of an RNA:DNA duplex.

Antisense compounds having a gapmer motif are considered chimeric antisense compounds. In a gapmer an internal region having a plurality of nucleotides that supports RNaseH cleavage is positioned between external regions having a plurality of nucleotides that are chemically distinct from the nucleosides of the internal region. In the case of an antisense oligonucleotide having a gapmer motif, the gap segment generally serves as the substrate for endonuclease cleavage, while the wing segments comprise modified nucleosides. In certain embodiments, the regions of a gapmer are differentiated by the types of sugar moieties comprising each distinct region. The types of sugar moieties that are used to differentiate the regions of a gapmer may in some embodiments include β-D-ribonucleosides, β-D-deoxyribonucleosides, 2′-modified nucleosides (such 2′-modified nucleosides may include 2′-MOE, and 2′-O—CH₃, among others), and bicyclic sugar modified nucleosides (such bicyclic sugar modified nucleosides may include those having a 4′-(CH₂)n-O-2′ bridge, where n=1 or n=2 and 4′-CH₂—O—CH₂-2′). In certain embodiments, wings may include several modified sugar moieties, including, for example 2′-MOE. In certain embodiments, wings may include several modified and unmodified sugar moieties. In certain embodiments, wings may include various combinations of 2′-MOE nucleosides and 2′-deoxynucleosides.

Each distinct region may comprise uniform sugar moieties, variant, or alternating sugar moieties. The wing-gap-wing motif is frequently described as “X—Y—Z”, where “X” represents the length of the 5′ wing, “Y” represents the length of the gap, and “Z” represents the length of the 3′ wing. “X” and “Z” may comprise uniform, variant, or alternating sugar moieties. In certain embodiments, “X” and “Y” may include one or more 2′-deoxynucleosides. “Y” may comprise 2′-deoxynucleosides. As used herein, a gapmer described as “X—Y—Z” has a configuration such that the gap is positioned immediately adjacent to each of the 5′ wing and the 3′ wing. Thus, no intervening nucleotides exist between the 5′ wing and gap, or the gap and the 3′ wing. Any of the antisense compounds described herein can have a gapmer motif. In certain embodiments, “X” and “Z” are the same; in other embodiments they are different.

In certain embodiments, gapmers provided herein include, for example 20-mers having a motif of 5-10-5.

In certain embodiments, gapmers provided herein include, for example 19-mers having a motif of 5-9-5.

In certain embodiments, gapmers provided herein include, for example 18-mers having a motif of 5-8-5.

In certain embodiments, gapmers provided herein include, for example 18-mers having a motif of 4-8-6.

In certain embodiments, gapmers provided herein include, for example 18-mers having a motif of 6-8-4.

In certain embodiments, gapmers provided herein include, for example 18-mers having a motif of 5-7-6.

Target Nucleic Acids, Target Regions and Nucleotide Sequences

Nucleotide sequences that encode Tau include, without limitation, the following: GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to U.S. Pat. No. 9,381,000 (incorporated herein as SEQ ID NO: 1), GENBANK Accession No. NM_001123066.3 (incorporated herein as SEQ ID NO: 2), GENBANK Accession No. NM_016841.4, a variant mRNA sequence which skips exons 3, 4, 6, 8, 10, and 12 (incorporated herein as SEQ ID NO: 3), GENBANK Accession No. NT_010783.14 truncated from nucleotides 2624000 to U.S. Pat. No. 2,761,000 (incorporated herein as SEQ ID NO: 4), GENBANK Accession No. DR002467.1 (incorporated herein as SEQ ID NO: 5), GENBANK Accession No. NM_001203251.1 (incorporated herein as SEQ ID NO: 6), and GENBANK Accession No. NM_016835.4 (incorporated herein as SEQ ID NO: 7).

It is understood that the sequence set forth in each SEQ ID NO in the Examples contained herein is independent of any modification to a sugar moiety, an internucleoside linkage, or a nucleobase. As such, antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an internucleoside linkage, or a nucleobase. Antisense compounds described by Isis Number (Isis No) indicate a combination of nucleobase sequence and motif.

In certain embodiments, a target region is a structurally defined region of the target nucleic acid. For example, a target region may encompass a 3′ UTR, a 5′ UTR, an exon, an intron, an exon/intron junction, a coding region, a translation initiation region, translation termination region, or other defined nucleic acid region. The structurally defined regions for Tau can be obtained by accession number from sequence databases such as NCBI and such information is incorporated herein by reference. In certain embodiments, a target region may encompass the sequence from a 5′ target site of one target segment within the target region to a 3′ target site of another target segment within the same target region.

Targeting includes determination of at least one target segment to which an antisense compound hybridizes, such that a desired effect occurs. In certain embodiments, the desired effect is a reduction in mRNA target nucleic acid levels. In certain embodiments, the desired effect is reduction of levels of protein encoded by the target nucleic acid or a phenotypic change associated with the target nucleic acid.

A target region may contain one or more target segments. Multiple target segments within a target region may be overlapping. Alternatively, they may be non-overlapping. In certain embodiments, target segments within a target region are separated by no more than about 300 nucleotides. In certain embodiments, target segments within a target region are separated by a number of nucleotides that is, is about, is no more than, is no more than about, 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 nucleotides on the target nucleic acid, or is a range defined by any two of the preceeding values. In certain embodiments, target segments within a target region are separated by no more than, or no more than about, 5 nucleotides on the target nucleic acid. In certain embodiments, target segments are contiguous. Contemplated are target regions defined by a range having a starting nucleic acid that is any of the 5′ target sites or 3′ target sites listed herein.

Suitable target segments may be found within a 5′ UTR, a coding region, a 3′ UTR, an intron, an exon, or an exon/intron junction. Target segments containing a start codon or a stop codon are also suitable target segments. A suitable target segment may specifically exclude a certain structurally defined region such as the start codon or stop codon.

The determination of suitable target segments may include a comparison of the sequence of a target nucleic acid to other sequences throughout the genome. For example, the BLAST algorithm may be used to identify regions of similarity amongst different nucleic acids. This comparison can prevent the selection of antisense compound sequences that may hybridize in a non-specific manner to sequences other than a selected target nucleic acid (i.e., non-target or off-target sequences).

There may be variation in activity (e.g., as defined by percent reduction of target nucleic acid levels) of the antisense compounds within an active target region. In certain embodiments, reductions in Tau mRNA levels are indicative of inhibition of Tau expression. Reductions in levels of a Tau protein are also indicative of inhibition of target mRNA expression. Reduction of percent of cells staining positive for hyperphosphorylated Tau are indicative of inhibition of Tau expression. Further, phenotypic changes are indicative of inhibition of Tau expression. Improvement in neurological function is indicative of inhibition of Tau expression. Improved memory and motor function are indicative of inhibition of Tau expression. Reduction of neurofibrillary inclusions is indicative of inhibition of Tau expression.

Hybridization

In some embodiments, hybridization occurs between an antisense compound disclosed herein and a Tau nucleic acid. The most common mechanism of hybridization involves hydrogen bonding (e.g., Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding) between complementary nucleobases of the nucleic acid molecules.

Hybridization can occur under varying conditions. Stringent conditions are sequence-dependent and are determined by the nature and composition of the nucleic acid molecules to be hybridized.

Methods of determining whether a sequence is specifically hybridizable to a target nucleic acid are well known in the art. In certain embodiments, the antisense compounds provided herein are specifically hybridizable with a Tau nucleic acid.

Complementarity

An antisense compound and a target nucleic acid are complementary to each other when a sufficient number of nucleobases of the antisense compound can hydrogen bond with the corresponding nucleobases of the target nucleic acid, such that a desired effect will occur (e.g., antisense inhibition of a target nucleic acid, such as a Tau nucleic acid).

Non-complementary nucleobases between an antisense compound and a Tau nucleic acid may be tolerated provided that the antisense compound remains able to specifically hybridize to a target nucleic acid. Moreover, an antisense compound may hybridize over one or more segments of a Tau nucleic acid such that intervening or adjacent segments are not involved in the hybridization event (e.g., a loop structure, mismatch or hairpin structure).

In certain embodiments, the antisense compounds provided herein, or a specified portion thereof, are, or are at least, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to a Tau nucleic acid, a target region, target segment, or specified portion thereof. Percent complementarity of an antisense compound with a target nucleic acid can be determined using routine methods.

For example, an antisense compound in which 18 of 20 nucleobases of the antisense compound are complementary to a target region, and would therefore specifically hybridize, would represent 90 percent complementarity. In this example, the remaining noncomplementary nucleobases may be clustered or interspersed with complementary nucleobases and need not be contiguous to each other or to complementary nucleobases. As such, an antisense compound which is 18 nucleobases in length having 4 (four) noncomplementary nucleobases which are flanked by two regions of complete complementarity with the target nucleic acid would have 77.8% overall complementarity with the target nucleic acid and would thus fall within the scope of the present invention. Percent complementarity of an antisense compound with a region of a target nucleic acid can be determined routinely using BLAST programs (basic local alignment search tools) and PowerBLAST programs known in the art (Altschul et al., J. Mol. Biol., 1990, 215, 403 410; Zhang and Madden, Genome Res., 1997, 7, 649 656). Percent homology, sequence identity or complementarity, can be determined by, for example, the Gap program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, Madison Wis.), using default settings, which uses the algorithm of Smith and Waterman (Adv. Appl. Math., 1981, 2, 482 489).

In certain embodiments, the antisense compounds provided herein, or specified portions thereof, are fully complementary (i.e., 100% complementary) to a target nucleic acid, or specified portion thereof. For example, an antisense compound may be fully complementary to a Tau nucleic acid, or a target region, or a target segment or target sequence thereof. As used herein, “fully complementary” means each nucleobase of an antisense compound is capable of precise base pairing with the corresponding nucleobases of a target nucleic acid. For example, a 20 nucleobase antisense compound is fully complementary to a target sequence that is 400 nucleobases long, so long as there is a corresponding 20 nucleobase portion of the target nucleic acid that is fully complementary to the antisense compound. Fully complementary can also be used in reference to a specified portion of the first and/or the second nucleic acid. For example, a 20 nucleobase portion of a 30 nucleobase antisense compound can be “fully complementary” to a target sequence that is 400 nucleobases long. The 20 nucleobase portion of the 30 nucleobase oligonucleotide is fully complementary to the target sequence if the target sequence has a corresponding 20 nucleobase portion wherein each nucleobase is complementary to the 20 nucleobase portion of the antisense compound. At the same time, the entire 30 nucleobase antisense compound may or may not be fully complementary to the target sequence, depending on whether the remaining 10 nucleobases of the antisense compound are also complementary to the target sequence.

The location of a non-complementary nucleobase may be at the 5′ end or 3′ end of the antisense compound. Alternatively, the non-complementary nucleobase or nucleobases may be at an internal position of the antisense compound. When two or more non-complementary nucleobases are present, they may be contiguous (i.e., linked) or non-contiguous. In one embodiment, a non-complementary nucleobase is located in the wing segment of a gapmer antisense oligonucleotide.

In certain embodiments, antisense compounds that are, or are up to 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleobases in length comprise no more than 4, no more than 3, no more than 2, or no more than 1 non-complementary nucleobase(s) relative to a target nucleic acid, such as a Tau nucleic acid, or specified portion thereof

In certain embodiments, antisense compounds that are, or are up to 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length comprise no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 non-complementary nucleobase(s) relative to a target nucleic acid, such as a Tau nucleic acid, or specified portion thereof

The antisense compounds provided herein also include those which are complementary to a portion of a target nucleic acid. As used herein, “portion” refers to a defined number of contiguous (i.e. linked) nucleobases within a region or segment of a target nucleic acid. A “portion” can also refer to a defined number of contiguous nucleobases of an antisense compound. In certain embodiments, the antisense compounds, are complementary to at least an 8 nucleobase portion of a target segment. In certain embodiments, the antisense compounds are complementary to at least a 9 nucleobase portion of a target segment. In certain embodiments, the antisense compounds are complementary to at least a 10 nucleobase portion of a target segment. In certain embodiments, the antisense compounds, are complementary to at least an 11 nucleobase portion of a target segment. In certain embodiments, the antisense compounds, are complementary to at least a 12 nucleobase portion of a target segment. In certain embodiments, the antisense compounds, are complementary to at least a 13 nucleobase portion of a target segment. In certain embodiments, the antisense compounds, are complementary to at least a 14 nucleobase portion of a target segment. In certain embodiments, the antisense compounds, are complementary to at least a 15 nucleobase portion of a target segment. Also contemplated are antisense compounds that are complementary to at least a 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more nucleobase portion of a target segment, or a range defined by any two of these values.

Identity

The antisense compounds provided herein may also have a defined percent identity to a particular nucleotide sequence, SEQ ID NO, or compound represented by a specific Isis number, or portion thereof. As used herein, an antisense compound is identical to the sequence disclosed herein if it has the same nucleobase pairing ability. For example, a RNA which contains uracil in place of thymidine in a disclosed DNA sequence would be considered identical to the DNA sequence since both uracil and thymidine pair with adenine. Shortened and lengthened versions of the antisense compounds described herein as well as compounds having non-identical bases relative to the antisense compounds provided herein also are contemplated. The non-identical bases may be adjacent to each other or dispersed throughout the antisense compound. Percent identity of an antisense compound is calculated according to the number of bases that have identical base pairing relative to the sequence to which it is being compared.

In certain embodiments, the antisense compounds, or portions thereof, are at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to one or more of the antisense compounds or SEQ ID NOs, or a portion thereof, disclosed herein.

In certain embodiments, a portion of the antisense compound is compared to an equal length portion of the target nucleic acid. In certain embodiments, an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleobase portion is compared to an equal length portion of the target nucleic acid.

In certain embodiments, a portion of the antisense oligonucleotide is compared to an equal length portion of the target nucleic acid. In certain embodiments, an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleobase portion is compared to an equal length portion of the target nucleic acid.

Modifications

A nucleoside is a base-sugar combination. The nucleobase (also known as base) portion of the nucleoside is normally a heterocyclic base moiety. Nucleotides are nucleosides that further include a phosphate group covalently linked to the sugar portion of the nucleoside. For those nucleosides that include a pentofuranosyl sugar, the phosphate group can be linked to the 2′, 3′ or 5′ hydroxyl moiety of the sugar. Oligonucleotides are formed through the covalent linkage of adjacent nucleosides to one another, to form a linear polymeric oligonucleotide. Within the oligonucleotide structure, the phosphate groups are commonly referred to as forming the internucleoside linkages of the oligonucleotide.

Modifications to antisense compounds encompass substitutions or changes to internucleoside linkages, sugar moieties, or nucleobases. Modified antisense compounds are often preferred over native forms because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for nucleic acid target, increased stability in the presence of nucleases, or increased inhibitory activity.

Chemically modified nucleosides may also be employed to increase the binding affinity of a shortened or truncated antisense oligonucleotide for its target nucleic acid. Consequently, comparable results can often be obtained with shorter antisense compounds that have such chemically modified nucleosides.

Modified Internucleoside Linkages

The naturally occurring internucleoside linkage of RNA and DNA is a 3′ to 5′ phosphodiester linkage. Antisense compounds having one or more modified, i.e. non-naturally occurring, internucleoside linkages are often selected over antisense compounds having naturally occurring internucleoside linkages because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for target nucleic acids, and increased stability in the presence of nucleases.

Oligonucleotides having modified internucleoside linkages include internucleoside linkages that retain a phosphorus atom as well as internucleoside linkages that do not have a phosphorus atom. Representative phosphorus containing internucleoside linkages include, but are not limited to, phosphodiesters, phosphotriesters, methylphosphonates, phosphoramidate, and phosphorothioates. Methods of preparation of phosphorous-containing and non-phosphorous-containing linkages are well known.

In certain embodiments, antisense compounds targeted to a Tau nucleic acid comprise one or more modified internucleoside linkages. In certain embodiments, the modified internucleoside linkages are interspersed throughout the antisense compound. In certain embodiments, the modified internucleoside linkages are phosphorothioate linkages. In certain embodiments, each internucleoside linkage of an antisense compound is a phosphorothioate internucleoside linkage.

Modified Sugar Moieties

Antisense compounds can optionally contain one or more nucleosides wherein the sugar group has been modified. Such sugar modified nucleosides may impart enhanced nuclease stability, increased binding affinity, or some other beneficial biological property to the antisense compounds. In certain embodiments, nucleosides comprise chemically modified ribofuranose ring moieties. Examples of chemically modified ribofuranose rings include without limitation, addition of substitutent groups (including 5′ and 2′ substituent groups, bridging of non-geminal ring atoms to form bicyclic nucleic acids (BNA), replacement of the ribosyl ring oxygen atom with S, N(R), or C(R₁)(R₂) (R, R₁ and R₂ are each independently H, C₁-C₁₂ alkyl or a protecting group) and combinations thereof. Examples of chemically modified sugars include 2′-F-5′-methyl substituted nucleoside (see PCT International Application WO 2008/101157 Published on Aug. 21, 2008 for other disclosed 5′,2′-bis substituted nucleosides) or replacement of the ribosyl ring oxygen atom with S with further substitution at the 2′-position (see published U.S. Patent Application US2005-0130923, published on Jun. 16, 2005) or alternatively 5′-substitution of a BNA (see PCT International Application WO 2007/134181 Published on Nov. 22, 2007 wherein LNA is substituted with for example a 5′-methyl or a 5′-vinyl group).

Examples of nucleosides having modified sugar moieties include without limitation nucleosides comprising 5′-vinyl, 5′-methyl (R or 5), 4′-S, 2′-F, 2′-OCH₃, 2′-OCH₂CH₃, 2′-OCH₂CH₂F and 2′-O(CH₂)₂OCH₃ substituent groups. The substituent at the 2′ position can also be selected from allyl, amino, azido, thio, O-allyl, O—C₁-C₁₀ alkyl, OCF₃, OCH₂F, O(CH₂)₂SCH₃, O(CH₂)₂—O—N(R_(m))(R_(n)), O—CH₂—C(═O)—N(R_(m))(R_(n)), and O—CH₂—C(═O)—N(R₁)—(CH₂)₂—N(R_(m))(R_(n)), where each R_(l), R_(m) and R_(n) is, independently, H or substituted or unsubstituted C₁-C₁₀ alkyl.

As used herein, “bicyclic nucleosides” refer to modified nucleosides comprising a bicyclic sugar moiety. Examples of bicyclic nucleosides include without limitation nucleosides comprising a bridge between the 4′ and the 2′ ribosyl ring atoms. In certain embodiments, antisense compounds provided herein include one or more bicyclic nucleosides comprising a 4′ to 2′ bridge. Examples of such 4′ to 2′ bridged bicyclic nucleosides, include but are not limited to one of the formulae: 4′-(CH₂)—O-2′ (LNA); 4′-(CH₂)—S-2; 4′-(CH₂)₂—O-2′ (ENA); 4′-CH(CH₃)—O-2′ and 4′-CH(CH₂OCH₃)—O-2′ (and analogs thereof see U.S. Pat. No. 7,399,845, issued on Jul. 15, 2008); 4′-C(CH₃)(CH₃)—O-2′ (and analogs thereof see published International Application WO/2009/006478, published Jan. 8, 2009); 4′-CH₂—N(OCH₃)-2′ (and analogs thereof see published International Application WO/2008/150729, published Dec. 11, 2008); 4′-CH₂—O—N(CH₃)-2′ (see published U.S. Patent Application US2004-0171570, published Sep. 2, 2004); 4′-CH₂—N(R)—O-2′, wherein R is H, C₁-C₁₂ alkyl, or a protecting group (see U.S. Pat. No. 7,427,672, issued on Sep. 23, 2008); 4′-CH₂—C(H)(CH₃)-2′ (see Chattopadhyaya et al., J. Org. Chem., 2009, 74, 118-134); and 4′-CH₂—C—(═CH₂)-2′ (and analogs thereof see published International Application WO 2008/154401, published on Dec. 8, 2008).

Further reports related to bicyclic nucleosides can also be found in published literature (see for example: Singh et al., Chem. Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54, 3607-3630; Wahlestedt et al., Proc. Natl. Acad. Sci. U.S.A, 2000, 97, 5633-5638; Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222; Singh et al., J. Org. Chem., 1998, 63, 10035-10039; Srivastava et al., J. Am. Chem. Soc., 2007, 129(26) 8362-8379; Elayadi et al., Curr. Opinion Invest. Drugs, 2001, 2, 558-561; Braasch et al., Chem. Biol., 2001, 8, 1-7; and Orum et al., Curr. Opinion Mol. Ther., 2001, 3, 239-243; U.S. Pat. Nos. 6,268,490; 6,525,191; 6,670,461; 6,770,748; 6,794,499; 7,034,133; 7,053,207; 7,399,845; 7,547,684; and 7,696,345; U.S. Patent Publication No. US2008-0039618; US2009-0012281; U.S. Patent Serial Nos. 60/989,574; 61/026,995; 61/026,998; 61/056,564; 61/086,231; 61/097,787; and 61/099,844; Published PCT International applications WO 1994/014226; WO 2004/106356; WO 2005/021570; WO 2007/134181; WO 2008/150729; WO 2008/154401; and WO 2009/006478. Each of the foregoing bicyclic nucleosides can be prepared having one or more stereochemical sugar configurations including for example α-L-ribofuranose and β-D-ribofuranose (see PCT international application PCT/DK98/00393, published on Mar. 25, 1999 as WO 99/14226).

In certain embodiments, bicyclic sugar moieties of BNA nucleosides include, but are not limited to, compounds having at least one bridge between the 4′ and the 2′ position of the pentofuranosyl sugar moiety wherein such bridges independently comprises 1 or from 2 to 4 linked groups independently selected from —[C(R_(a))(R_(b))]_(n)—, —C(R_(a))═C(R_(b))—, —C(R_(a))═N—, —C(═O)—, —C(═NR_(a))—, —C(═S)—, —O—, —Si(R_(a))₂—, —S(═O)_(x)—, and —N(R_(a))—;

wherein:

x is 0, 1, or 2;

n is 1, 2, 3, or 4;

each R_(a) and R_(b) is, independently, H, a protecting group, hydroxyl, C₁-C₁₂ alkyl, substituted C₁-C₁₂ alkyl, C₂-C₁₂ alkenyl, substituted C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, substituted C₂-C₁₂ alkynyl, C₅-C₂₀ aryl, substituted C₅-C₂₀ aryl, heterocycle radical, substituted heterocycle radical, heteroaryl, substituted heteroaryl, C₅-C₇ alicyclic radical, substituted C₅-C₇ alicyclic radical, halogen, OJ₁, NJ₁J₂, SJ₁, N₃, COOJ₁, acyl (C(═O)—H), substituted acyl, CN, sulfonyl (S(═O)₂-J₁), or sulfoxyl (S(═O)-J₁); and

each J₁ and J₂ is, independently, H, C₁-C₁₂ alkyl, substituted C₁-C₁₂ alkyl, C₂-C₁₂ alkenyl, substituted C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, substituted C₂-C₁₂ alkynyl, C₅-C₂₀ aryl, substituted C₅-C₂₀ aryl, acyl (C(═O)—H), substituted acyl, a heterocycle radical, a substituted heterocycle radical, C₁-C₁₂ aminoalkyl, substituted C₁-C₁₂ aminoalkyl or a protecting group.

In certain embodiments, the bridge of a bicyclic sugar moiety is —[C(R_(a))(R_(b))]_(n)—, —[C(R_(a))(R_(b))]_(n)—O—, —C(R_(a)R_(b))—N(R)—O— or —C(R_(a)R_(b))—O—N(R)—. In certain embodiments, the bridge is 4′-CH₂-2′, 4′-(CH₂)₂-2′, 4′-(CH₂)₃-2′, 4′-CH₂—O-2′, 4′-(CH₂)₂—O-2′, 4′-CH₂—O—N(R)-2′ and 4′-CH₂—N(R)—O-2′- wherein each R is, independently, H, a protecting group or C₁-C₁₂ alkyl.

In certain embodiments, bicyclic nucleosides are further defined by isomeric configuration. For example, a nucleoside comprising a 4′-2′ methylene-oxy bridge, may be in the α-L configuration or in the β-D configuration. Previously, α-L-methyleneoxy (4′-CH₂—O-2′) BNA's have been incorporated into antisense oligonucleotides that showed antisense activity (Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372).

In certain embodiments, bicyclic nucleosides include, but are not limited to, (A) α-L-methyleneoxy (4′-CH₂—O-2′) BNA, (B) β-D-methyleneoxy (4′-CH₂—O-2′) BNA, (C) ethyleneoxy (4′-(CH₂)₂—O-2′) BNA, (D) aminooxy (4′-CH₂—O—N(R)-2′) BNA, (E) oxyamino (4′-CH₂—N(R)—O-2′) BNA, and (F) methyl(methyleneoxy) (4′-CH(CH₃)—O-2′) BNA, (G) methylene-thio (4′-CH₂—S-2′) BNA, (H) methylene-amino (4′-CH₂—N(R)-2′) BNA, (I) methyl carbocyclic (4′-CH₂—CH(CH₃)-2′) BNA, and (J) propylene carbocyclic (4′-(CH₂)₃-2′) BNA as depicted below.

wherein Bx is the base moiety and R is independently H, a protecting group or C₁-C₁₂ alkyl.

In certain embodiments, bicyclic nucleosides are provided having Formula I:

wherein:

Bx is a heterocyclic base moiety;

-Q_(a)-Q_(b)-Q_(c)- is —CH₂—N(R_(c))—CH₂—, —C(═O)—N(R_(c))—CH₂—, —CH₂—O—N(R_(c))—, —CH₂—N(R_(c))—O— or —N(R_(c))—O—CH₂;

R_(c) is C₁-C₁₂ alkyl or an amino protecting group; and

T_(a) and T_(b) are each, independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety or a covalent attachment to a support medium.

In certain embodiments, bicyclic nucleosides are provided having Formula II:

wherein:

Bx is a heterocyclic base moiety;

T_(a) and T_(b) are each, independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety or a covalent attachment to a support medium;

Z_(a) is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, substituted C₁-C₆ alkyl, substituted C₂-C₆ alkenyl, substituted C₂-C₆ alkynyl, acyl, substituted acyl, substituted amide, thiol or substituted thio.

In one embodiment, each of the substituted groups is, independently, mono or poly substituted with substituent groups independently selected from halogen, oxo, hydroxyl, OJ_(c), NJ_(c)J_(d), SJ_(c), N₃, OC(═X)J_(c), and NJ_(c)C(═X)NJ_(c)J_(d), wherein each J_(c), J_(d) and J_(c) is, independently, H, C₁-C₆ alkyl, or substituted C₁-C₆ alkyl and X is O or NJ_(c).

In certain embodiments, bicyclic nucleosides are provided having Formula III:

wherein:

Bx is a heterocyclic base moiety;

T_(a) and T_(b) are each, independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety or a covalent attachment to a support medium;

Z_(b) is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, substituted C₁-C₆ alkyl, substituted C₂-C₆ alkenyl, substituted C₂-C₆ alkynyl or substituted acyl (C(═O)—).

In certain embodiments, bicyclic nucleosides are provided having Formula IV:

wherein:

Bx is a heterocyclic base moiety;

T_(a) and T_(b) are each, independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety or a covalent attachment to a support medium;

R_(d) is C₁-C₆ alkyl, substituted C₁-C₆ alkyl, C₂-C₆ alkenyl, substituted C₂-C₆ alkenyl, C₂-C₆ alkynyl or substituted C₂-C₆ alkynyl;

each q_(a), q_(b), q_(c) and q_(d) is, independently, H, halogen, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, C₂-C₆ alkenyl, substituted C₂-C₆ alkenyl, C₂-C₆ alkynyl or substituted C₂-C₆ alkynyl, C₁-C₆ alkoxyl, substituted C₁-C₆ alkoxyl, acyl, substituted acyl, C₁-C₆ aminoalkyl or substituted C₁-C₆ aminoalkyl;

In certain embodiments, bicyclic nucleosides are provided having Formula V:

wherein:

Bx is a heterocyclic base moiety;

T_(a) and T_(b) are each, independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety or a covalent attachment to a support medium;

q_(a), q_(b), q_(e) and q_(f) are each, independently, hydrogen, halogen, C₁-C₁₂ alkyl, substituted C₁-C₁₂ alkyl, C₂-C₁₂ alkenyl, substituted C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, substituted C₂-C₁₂ alkynyl, C₁-C₁₂ alkoxy, substituted C₁-C₁₂ alkoxy, OJ_(j), SJ_(j), SOJ_(j), SO₂J_(j), NJ_(j)J_(k), N₃, CN, C(═O)OJ_(j), C(═O)NJ_(j)J_(k), C(═O)J_(j), O—C(═O)NJ_(j)J_(k), N(H)C(═NH)NJ_(j)J_(k), N(H)C(═O)NJ_(j)J_(k) or N(H)C(═S)NJ_(j)J_(k);

or q_(e) and q_(f) together are ═C(q_(g))(q_(h));

q_(g) and q_(h) are each, independently, H, halogen, C₁-C₁₂ alkyl or substituted C₁-C₁₂ alkyl.

The synthesis and preparation of the methyleneoxy (4′-CH₂—O-2′) BNA monomers adenine, cytosine, guanine, 5-methyl-cytosine, thymine and uracil, along with their oligomerization, and nucleic acid recognition properties have been described (Koshkin et al., Tetrahedron, 1998, 54, 3607-3630). BNAs and preparation thereof are also described in WO 98/39352 and WO 99/14226.

Analogs of methyleneoxy (4′-CH₂—O-2′) BNA and 2′-thio-BNAs, have also been prepared (Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222). Preparation of locked nucleoside analogs comprising oligodeoxyribonucleotide duplexes as substrates for nucleic acid polymerases has also been described (Wengel et al., WO 99/14226). Furthermore, synthesis of 2′-amino-BNA, a novel comformationally restricted high-affinity oligonucleotide analog has been described in the art (Singh et al., J. Org. Chem., 1998, 63, 10035-10039). In addition, 2′-amino- and 2′-methylamino-BNA's have been prepared and the thermal stability of their duplexes with complementary RNA and DNA strands has been previously reported.

In certain embodiments, bicyclic nucleosides are provided having Formula VI:

wherein:

Bx is a heterocyclic base moiety;

T_(a) and T_(b) are each, independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety or a covalent attachment to a support medium;

each q_(i), q_(j), q_(k) and q_(l) is, independently, H, halogen, C₁-C₁₂ alkyl, substituted C₁-C₁₂ alkyl, C₂-C₁₂ alkenyl, substituted C₂-C₁₂ alkenyl, C₂-C₁₂ alkynyl, substituted C₂-C₁₂ alkynyl, C₁-C₁₂ alkoxyl, substituted C₁-C₁₂ alkoxyl, OJ_(j), SJ_(j), SOJ_(j), SO₂J_(j), NJ_(j)J_(k), N₃, CN, C(═O)OJ_(j), C(═O)NJ_(j)J_(k), C(═O)J_(j), O—C(═O)NJ_(j)J_(k), N(H)C(═NH)NJ_(j)J_(k), N(H)C(═O)NJ_(j)J_(k) or N(H)C(═S)NJ_(j)J_(k); and

q_(i) and q_(j) or q_(l) and q_(k) together are ═C(q_(g))(q_(h)), wherein q_(g) and q_(h) are each, independently, H, halogen, C₁-C₁₂ alkyl or substituted C₁-C₁₂ alkyl.

One carbocyclic bicyclic nucleoside having a 4′-(CH₂)₃-2′ bridge and the alkenyl analog bridge 4′-CH═CH—CH₂-2′ have been described (Freier et al., Nucleic Acids Research, 1997, 25(22), 4429-4443 and Albaek et al., J. Org. Chem., 2006, 71, 7731-7740). The synthesis and preparation of carbocyclic bicyclic nucleosides along with their oligomerization and biochemical studies have also been described (Srivastava et al., J. Am. Chem. Soc., 2007, 129(26), 8362-8379).

As used herein, “4′-2′ bicyclic nucleoside” or “4′ to 2′ bicyclic nucleoside” refers to a bicyclic nucleoside comprising a furanose ring comprising a bridge connecting two carbon atoms of the furanose ring connects the 2′ carbon atom and the 4′ carbon atom of the sugar ring.

As used herein, “monocylic nucleosides” refer to nucleosides comprising modified sugar moieties that are not bicyclic sugar moieties. In certain embodiments, the sugar moiety, or sugar moiety analogue, of a nucleoside may be modified or substituted at any position.

As used herein, “2′-modified sugar” means a furanosyl sugar modified at the 2′ position. In certain embodiments, such modifications include substituents selected from: a halide, including, but not limited to substituted and unsubstituted alkoxy, substituted and unsubstituted thioalkyl, substituted and unsubstituted amino alkyl, substituted and unsubstituted alkyl, substituted and unsubstituted allyl, and substituted and unsubstituted alkynyl. In certain embodiments, 2′ modifications are selected from substituents including, but not limited to: O[(CH₂)_(n)O]_(m)CH₃, O(CH₂)_(n)NH₂, O(CH₂)_(n)CH₃, O(CH₂)_(n)F, O(CH₂)—ONH₂, OCH₂C(═O)N(H)CH₃, and O(CH₂)_(n)ON[(CH₂)_(n)CH₃]₂, where n and m are from 1 to about 10. Other 2′-substituent groups can also be selected from: C₁-C₁₂ alkyl, substituted alkyl, alkenyl, alkynyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH₃, OCN, Cl, Br, CN, F, CF₃, OCF₃, SOCH₃, SO₂CH₃, ONO₂, NO₂, N₃, NH₂, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, an RNA cleaving group, a reporter group, an intercalator, a group for improving pharmacokinetic properties, or a group for improving the pharmacodynamic properties of an antisense compound, and other substituents having similar properties. In certain embodiments, modified nucleosides comprise a 2′-MOE side chain (Baker et al., J. Biol. Chem., 1997, 272, 11944-12000). Such 2′-MOE substitution have been described as having improved binding affinity compared to unmodified nucleosides and to other modified nucleosides, such as 2′-O-methyl, O-propyl, and O-aminopropyl. Oligonucleotides having the 2′-MOE substituent also have been shown to be antisense inhibitors of gene expression with promising features for in vivo use (Martin, Helv. Chim. Acta, 1995, 78, 486-504; Altmann et al., Chimia, 1996, 50, 168-176; Altmann et al., Biochem. Soc. Trans., 1996, 24, 630-637; and Altmann et al., Nucleosides Nucleotides, 1997, 16, 917-926).

As used herein, a “modified tetrahydropyran nucleoside” or “modified THP nucleoside” means a nucleoside having a six-membered tetrahydropyran “sugar” substituted in for the pentofuranosyl residue in normal nucleosides (a sugar surrogate). Modified THP nucleosides include, but are not limited to, what is referred to in the art as hexitol nucleic acid (HNA), anitol nucleic acid (ANA), manitol nucleic acid (MNA) (see Leumann, Bioorg. Med. Chem., 2002, 10, 841-854), fluoro HNA (F-HNA) or those compounds having Formula VII:

wherein independently for each of said at least one tetrahydropyran nucleoside analog of Formula VII:

Bx is a heterocyclic base moiety;

T_(a) and T_(b) are each, independently, an internucleoside linking group linking the tetrahydropyran nucleoside analog to the antisense compound or one of T_(a) and T_(b) is an internucleoside linking group linking the tetrahydropyran nucleoside analog to the antisense compound and the other of T_(a) and T_(b) is H, a hydroxyl protecting group, a linked conjugate group or a 5′ or 3′-terminal group;

q₁, q₂, q₃, q₄, q₅, q₆ and q₇ are each independently, H, C₁-C₆ alkyl, substituted C₁-C₆ alkyl, C₂-C₆ alkenyl, substituted C₂-C₆ alkenyl, C₂-C₆ alkynyl or substituted C₂-C₆ alkynyl; and each of R₁ and R₂ is selected from hydrogen, hydroxyl, halogen, substituted or unsubstituted alkoxy, NJ₁J₂, SJ₁, N₃, OC(═X)J₁, OC(═X)NJ₁J₂, NJ₃C(═X)NJ₁J₂ and CN, wherein X is O, S or NJ₁ and each J₁, J₂ and J₃ is, independently, H or C₁-C₆ alkyl.

In certain embodiments, the modified THP nucleosides of Formula VII are provided wherein q₁, q₂, q₃, q₄, q₅, q₆ and q₇ are each H. In certain embodiments, at least one of q₁, q₂, q₃, q₄, q₅, q₆ and q₇ is other than H. In certain embodiments, at least one of q₁, q₂, q₃, q₄, q₅, q₆ and q₇ is methyl. In certain embodiments, THP nucleosides of Formula VII are provided wherein one of R₁ and R₂ is fluoro. In certain embodiments, R₁ is fluoro and R₂ is H; R₁ is methoxy and R₂ is H, and R₁ is H and R₂ is methoxyethoxy.

As used herein, “2′-modified” or “2′-substituted” refers to a nucleoside comprising a sugar comprising a substituent at the 2′ position other than H or OH. 2′-modified nucleosides, include, but are not limited to, bicyclic nucleosides wherein the bridge connecting two carbon atoms of the sugar ring connects the 2′ carbon and another carbon of the sugar ring; and nucleosides with non-bridging 2′ substituents, such as allyl, amino, azido, thio, O-allyl, O—C₁-C₁₀ alkyl, —OCF₃, O—(CH₂)₂—O—CH₃, 2′-O(CH₂)₂SCH₃, O—(CH₂)₂—O—N(R_(m))(R_(n)), or O—CH₂—C(═O)—N(R_(m))(R_(n)), where each R_(m) and R_(n) is, independently, H or substituted or unsubstituted C₁-C₁₀ alkyl. 2′-modifed nucleosides may further comprise other modifications, for example at other positions of the sugar and/or at the nucleobase.

As used herein, “2′-F” refers to a nucleoside comprising a sugar comprising a fluoro group at the 2′ position.

As used herein, “2′-OMe” or “2′-OCH₃” or “2′-O-methyl” each refers to a nucleoside comprising a sugar comprising an —OCH₃ group at the 2′ position of the sugar ring.

As used herein, “MOE” or “2′-MOE” or “2′-OCH₂CH₂OCH₃” or “2′-O-methoxyethyl” each refers to a nucleoside comprising a sugar comprising a —OCH₂CH₂OCH₃ group at the 2′ position of the sugar ring.

As used herein, “oligonucleotide” refers to a compound comprising a plurality of linked nucleosides. In certain embodiments, one or more of the plurality of nucleosides is modified. In certain embodiments, an oligonucleotide comprises one or more ribonucleosides (RNA) and/or deoxyribonucleosides (DNA).

Many other bicyclo and tricyclo sugar surrogate ring systems are also known in the art that can be used to modify nucleosides for incorporation into antisense compounds (see for example review article: Leumann, Bioorg. Med. Chem., 2002, 10, 841-854).

Such ring systems can undergo various additional substitutions to enhance activity.

Methods for the preparations of modified sugars are well known to those skilled in the art.

In nucleotides having modified sugar moieties, the nucleobase moieties (natural, modified or a combination thereof) are maintained for hybridization with an appropriate nucleic acid target.

In certain embodiments, antisense compounds comprise one or more nucleosides having modified sugar moieties. In certain embodiments, the modified sugar moiety is 2′-MOE. In certain embodiments, the 2′-MOE modified nucleosides are arranged in a gapmer motif. In certain embodiments, the modified sugar moiety is a bicyclic nucleoside having a (4′-CH(CH₃)—O-2′) bridging group. In certain embodiments, the (4′-CH(CH₃)—O-2′) modified nucleosides are arranged throughout the wings of a gapmer motif

Compositions and Methods for Formulating Pharmaceutical Compositions

Antisense oligonucleotides may be admixed with pharmaceutically acceptable active or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions are dependent upon a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.

An antisense compound targeted to a Tau nucleic acid can be utilized in pharmaceutical compositions by combining the antisense compound with a suitable pharmaceutically acceptable diluent or carrier. A pharmaceutically acceptable diluent includes phosphate-buffered saline (PBS). PBS is a diluent suitable for use in compositions to be delivered parenterally. Accordingly, in one embodiment, employed in the methods described herein is a pharmaceutical composition comprising an antisense compound targeted to a Tau nucleic acid and a pharmaceutically acceptable diluent. In certain embodiments, the pharmaceutically acceptable diluent is PBS. In certain embodiments, the antisense compound is an antisense oligonucleotide.

Pharmaceutical compositions comprising antisense compounds encompass any pharmaceutically acceptable salts, esters, or salts of such esters, or any other oligonucleotide which, upon administration to an animal, including a human, is capable of providing (directly or indirectly) the biologically active metabolite or residue thereof. Accordingly, for example, the disclosure is also drawn to pharmaceutically acceptable salts of antisense compounds, prodrugs, pharmaceutically acceptable salts of such prodrugs, and other bioequivalents. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.

A prodrug can include the incorporation of additional nucleosides at one or both ends of an antisense compound which are cleaved by endogenous nucleases within the body, to form the active antisense compound.

Conjugated Antisense Compounds

Antisense compounds may be covalently linked to one or more moieties or conjugates which enhance the activity, cellular distribution or cellular uptake of the resulting antisense oligonucleotides. Typical conjugate groups include cholesterol moieties and lipid moieties. Additional conjugate groups include carbohydrates, phospholipids, biotin, phenazine, folate, phenanthridine, anthraquinone, acridine, fluoresceins, rhodamines, coumarins, and dyes.

Antisense compounds can also be modified to have one or more stabilizing groups that are generally attached to one or both termini of antisense compounds to enhance properties such as, for example, nuclease stability. Included in stabilizing groups are cap structures. These terminal modifications protect the antisense compound having terminal nucleic acid from exonuclease degradation, and can help in delivery and/or localization within a cell. The cap can be present at the 5′-terminus (5′-cap), or at the 3′-terminus (3′-cap), or can be present on both termini. Cap structures are well known in the art and include, for example, inverted deoxy abasic caps. Further 3′ and 5′-stabilizing groups that can be used to cap one or both ends of an antisense compound to impart nuclease stability include those disclosed in WO 03/004602 published on Jan. 16, 2003.

Cell Culture and Antisense Compounds Treatment

The effects of antisense compounds on the level, activity or expression of Tau nucleic acids can be tested in vitro in a variety of cell types. Cell types used for such analyses are available from commercial vendors (e.g. American Type Culture Collection, Manassas, Va.; Zen-Bio, Inc., Research Triangle Park, N.C.; Clonetics Corporation, Walkersville, Md.) and are cultured according to the vendor's instructions using commercially available reagents (e.g. Invitrogen Life Technologies, Carlsbad, Calif.). Illustrative cell types include, but are not limited to, HepG2 cells, Hep3B cells, and primary hepatocytes.

In Vitro Testing of Antisense Oligonucleotides

Described herein are methods for treatment of cells with antisense oligonucleotides, which can be modified appropriately for treatment with other antisense compounds.

Cells may be treated with antisense oligonucleotides when the cells reach approximately 60-80% confluency in culture.

One reagent commonly used to introduce antisense oligonucleotides into cultured cells includes the cationic lipid transfection reagent LIPOFECTIN (Invitrogen, Carlsbad, Calif.). Antisense oligonucleotides may be mixed with LIPOFECTIN in OPTI-MEM 1 (Invitrogen, Carlsbad, Calif.) to achieve the desired final concentration of antisense oligonucleotide and a LIPOFECTIN concentration that may range from 2 to 12 ug/mL per 100 nM antisense oligonucleotide.

Another reagent used to introduce antisense oligonucleotides into cultured cells includes LIPOFECTAMINE (Invitrogen, Carlsbad, Calif.). Antisense oligonucleotide is mixed with LIPOFECTAMINE in OPTI-MEM 1 reduced serum medium (Invitrogen, Carlsbad, Calif.) to achieve the desired concentration of antisense oligonucleotide and a LIPOFECTAMINE concentration that may range from 2 to 12 ug/mL per 100 nM antisense oligonucleotide.

Another technique used to introduce antisense oligonucleotides into cultured cells includes electroporation.

Cells are treated with antisense oligonucleotides by routine methods. Cells may be harvested 16-24 hours after antisense oligonucleotide treatment, at which time RNA or protein levels of target nucleic acids are measured by methods known in the art and described herein. In general, when treatments are performed in multiple replicates, the data are presented as the average of the replicate treatments.

The concentration of antisense oligonucleotide used varies from cell line to cell line. Methods to determine the optimal antisense oligonucleotide concentration for a particular cell line are well known in the art. Antisense oligonucleotides are typically used at concentrations ranging from 1 nM to 300 nM when transfected with LIPOFECTAMINE. Antisense oligonucleotides are used at higher concentrations ranging from 625 to 20,000 nM when transfected using electroporation.

RNA Isolation

RNA analysis can be performed on total cellular RNA or poly(A)+ mRNA. Methods of RNA isolation are well known in the art. RNA is prepared using methods well known in the art, for example, using the TRIZOL Reagent (Invitrogen, Carlsbad, Calif.) according to the manufacturer's recommended protocols.

Analysis of Inhibition of Target Levels or Expression

Inhibition of levels or expression of a Tau nucleic acid can be assayed in a variety of ways known in the art. For example, target nucleic acid levels can be quantitated by, e.g., Northern blot analysis, competitive polymerase chain reaction (PCR), or quantitative real-time PCR. RNA analysis can be performed on total cellular RNA or poly(A)+ mRNA. Methods of RNA isolation are well known in the art. Northern blot analysis is also routine in the art. Quantitative real-time PCR can be conveniently accomplished using the commercially available ABI PRISM 7600, 7700, or 7900 Sequence Detection System, available from PE-Applied Biosystems, Foster City, Calif. and used according to manufacturer's instructions.

Quantitative Real-Time PCR Analysis of Target RNA Levels

Quantitation of target RNA levels may be accomplished by quantitative real-time PCR using the ABI PRISM 7600, 7700, or 7900 Sequence Detection System (PE-Applied Biosystems, Foster City, Calif.) according to manufacturer's instructions. Methods of quantitative real-time PCR are well known in the art.

Prior to real-time PCR, the isolated RNA is subjected to a reverse transcriptase (RT) reaction, which produces complementary DNA (cDNA) that is then used as the substrate for the real-time PCR amplification. The RT and real-time PCR reactions are performed sequentially in the same sample well. RT and real-time PCR reagents may be obtained from Invitrogen (Carlsbad, Calif.). RT real-time-PCR reactions are carried out by methods well known to those skilled in the art.

Gene (or RNA) target quantities obtained by real time PCR are normalized using either the expression level of a gene whose expression is constant, such as cyclophilin A, or by quantifying total RNA using RIBOGREEN (Invitrogen, Inc. Carlsbad, Calif.). Cyclophilin A expression is quantified by real time PCR, by being run simultaneously with the target, multiplexing, or separately. Total RNA is quantified using RIBOGREEN RNA quantification reagent (Invetrogen, Inc. Eugene, Oreg.). Methods of RNA quantification by RIBOGREEN are taught in Jones, L. J., et al, (Analytical Biochemistry, 1998, 265, 368-374). A CYTOFLUOR 4000 instrument (PE Applied Biosystems) is used to measure RIBOGREEN fluorescence.

Probes and primers are designed to hybridize to a Tau nucleic acid. Methods for designing real-time PCR probes and primers are well known in the art, and may include the use of software such as PRIMER EXPRESS Software (Applied Biosystems, Foster City, Calif.).

Analysis of Protein Levels

Antisense inhibition of Tau nucleic acids can be assessed by measuring Tau protein levels. Protein levels of Tau can be evaluated or quantitated in a variety of ways well known in the art, such as immunoprecipitation, Western blot analysis (immunoblotting), enzyme-linked immunosorbent assay (ELISA), quantitative protein assays, protein activity assays (for example, caspase activity assays), immunohistochemistry, immunocytochemistry or fluorescence-activated cell sorting (FACS). Antibodies directed to a target can be identified and obtained from a variety of sources, such as the MSRS catalog of antibodies (Aerie Corporation, Birmingham, Mich.), or can be prepared via conventional monoclonal or polyclonal antibody generation methods well known in the art.

In Vivo Testing of Antisense Compounds

Antisense compounds, for example, antisense oligonucleotides, are tested in animals to assess their ability to inhibit expression of Tau and produce phenotypic changes, such as, improved cognition and motor function. In certain embodiments, cognition is measured by novel object recognition and nestlet building activity. In certain embodiments, motor function is measured by walking initiation analysis, rotarod, grip strength, pole climb, open field performance, balance beam, hindpaw footprint testing in the animal. In certain embodiments, antisense compounds, for example, antisense oligonucleotides, are tested in animals to assess their ability to reduce hyperphosphorylated tau and neurofibrillary tangles. In certain embodiments, antisense compounds, for example, antisense oligonucleotides, are tested to assess their ability to prevent, and/or reduce severity of, seizures in a pentylenetetrazol (PTZ) induced seizure model.

Testing may be performed in normal animals, or in experimental disease models. For administration to animals, antisense oligonucleotides are formulated in a pharmaceutically acceptable diluent, such as phosphate-buffered saline. Administration includes parenteral routes of administration, such as intraperitoneal, intravenous, and subcutaneous. Calculation of antisense oligonucleotide dosage and dosing frequency is within the abilities of those skilled in the art, and depends upon factors such as route of administration and animal body weight. Following a period of treatment with antisense oligonucleotides, RNA is isolated from CNS tissue or CSF and changes in Tau nucleic acid expression are measured.

Certain Indications

In certain embodiments, provided herein are methods, compounds, and compositions of treating an individual comprising administering one or more pharmaceutical compositions described herein. In certain embodiments, the individual has a neurodegenerative disease. In certain embodiments, the individual is at risk for developing a neurodegenerative disease, including, but not limited to, a Tauopathy, Alzheimer's Disease, Fronto-temporal Dementia (FTD), FTDP-17, Progressive Supranuclear Palsy (PSP), Chronic Traumatic Encephalopathy (CTE), Corticobasal Ganglionic Degeneration (CBD), Epilepsy, and Dravet's Syndrome. In certain embodiments, the individual has been identified as having a Tau associated disease. In certain embodiments, provided herein are methods for prophylactically reducing Tau expression in an individual. Certain embodiments include treating an individual in need thereof by administering to an individual a therapeutically effective amount of an antisense compound targeted to a Tau nucleic acid.

In one embodiment, administration of a therapeutically effective amount of an antisense compound targeted to a Tau nucleic acid is accompanied by monitoring of Tau levels in an individual, to determine an individual's response to administration of the antisense compound. An individual's response to administration of the antisense compound may be used by a physician to determine the amount and duration of therapeutic intervention.

In certain embodiments, administration of an antisense compound targeted to a Tau nucleic acid results in reduction of Tau expression by at least 15, 20, 25, 30, 35, 40, 45, 50, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100%, or a range defined by any two of these values. In certain embodiments, administration of an antisense compound targeted to a Tau nucleic acid results in improved motor function in an animal. In certain embodiments, administration of a Tau antisense compound improves motor function by at least 15, 20, 25, 30, 35, 40, 45, 50, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100%, or a range defined by any two of these values.

In certain embodiments, pharmaceutical compositions comprising an antisense compound targeted to Tau are used for the preparation of a medicament for treating a patient suffering or susceptible to a neurodegenerative disease including a Tauopathy, Alzheimer's Disease, Fronto-temporal Dementia (FTD), FTDP-17, Progressive Supranuclear Palsy (PSP), Chronic Traumatic Encephalopathy (CTE), Corticobasal Ganglionic Degeneration (CBD), Epilepsy, and Dravet's Syndrome.

Certain Hotspot Regions

1. Nucleobases 135783-135980 of SEQ ID NO: 1

In certain embodiments, antisense oligonucleotides are designed to target nucleobases 135783-135980 of SEQ ID NO: 1 (GENBANK Accession No. GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to 9381000). In certain embodiments, nucleobases 135783-135980 are a hotspot region. In certain embodiments, nucleobases 135783-135980 are targeted by antisense oligonucleotides. In certain embodiments, the antisense oligonucleotides are 18, 19, or 20 nucleobases in length. In certain embodiments, the antisense oligonucleotides are gapmers. In certain embodiments, the gapmers are 5-10-5 MOE gapmers, 5-9-5 MOE gapmers, 5-7-6 MOE gapmers, and 5-8-5 MOE gapmers. In certain embodiments, the nucleosides of the antisense oligonucleotides are linked by phosphorothioate internucleoside linkages. In certain embodiments, the nucleosides of the antisense oligonucleotides are linked by phosphodiester internucleoside linkages. In certain embodiments, the nucleosides of the antisense oligonucleotides are linked by phosphorothioate and phosphodiester internucleotide linkages (e.g., the antisense oligonucleotides have “mixed backbones”).

In certain embodiments, nucleobases 135783-135980 are targeted by the following ISIS numbers: 424879, 424880, 548937, 613114-613120, 622096-622150, 623988-623996, 664511-664542, and 664661-664819.

In certain embodiments, nucleobases 135783-135980 are targeted by the following SEQ ID NOs: 56, 57, 248, 462-467, 1668-1698, 2025-2048, 2301-2309, 2331-2443, and 2478-2483.

In certain embodiments, antisense oligonucleotides targeting nucleobases 135783-135980 achieve at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31%, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41%, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, or at least 93% reduction of Tau mRNA and/or protein levels in vitro and/or in vivo.

2. Nucleobases 135853-135872 of SEQ ID NO: 1

In certain embodiments, antisense oligonucleotides are designed to target nucleobases 135853-135872 of SEQ ID NO: 1 (GENBANK Accession No. GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to 9381000). In certain embodiments, nucleobases 135853-135872 are a hotspot region. In certain embodiments, nucleobases 135853-135872 are targeted by antisense oligonucleotides. In certain embodiments, the antisense oligonucleotides are 18, 19, or 20 nucleobases in length. In certain embodiments, the antisense oligonucleotides are gapmers. In certain embodiments, the gapmers are 5-10-5 MOE gapmers, 5-9-5 MOE gapmers, 5-7-6 MOE gapmers, or 5-8-5 MOE gapmers. In certain embodiments, the nucleosides of the antisense oligonucleotides are linked by phosphorothioate internucleoside linkages. In certain embodiments, the nucleosides of the antisense oligonucleotides are linked by phosphodiester internucleoside linkages. In certain embodiments, the nucleosides of the antisense oligonucleotides are linked by phosphorothioate and phosphodiester internucleotide linkages (e.g., the antisense oligonucleotides have “mixed backbones”).

In certain embodiments, nucleobases 135853-135872 are targeted by the following ISIS numbers: 424879, 424880, 613117, 613118, 622114-622125, 623993-623996, 664522-664542, 664676-664713, 664729-664766, and 664783-664819.

In certain embodiments, nucleobases 135853-135872 are targeted by the following SEQ ID NOs: 56, 57, 248, 464-465, 1668-1673, 2039-2048, 2306-2309, 2345-2443, and 2478-2483.

In certain embodiments, antisense oligonucleotides targeting nucleobases 135853-135872 achieve at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41%, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, or at least 87% reduction of Tau mRNA and/or protein levels in vitro and/or in vivo.

3. Nucleobases 135783-135929 of SEQ ID NO: 1

In certain embodiments, antisense oligonucleotides are designed to target nucleobases 135783-135929 of SEQ ID NO: 1 (GENBANK Accession No. GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to 9381000). In certain embodiments, nucleobases 135783-135929 are a hotspot region. In certain embodiments, nucleobases 135783-135929 are targeted by antisense oligonucleotides. In certain embodiments, the antisense oligonucleotides are 18, 19, or 20 nucleobases in length. In certain embodiments, the antisense oligonucleotides are gapmers. In certain embodiments, the gapmers are 5-10-5 MOE gapmers, 5-9-5 MOE gapmers, 5-7-6 MOE gapmers, or 5-8-5 MOE gapmers. In certain embodiments, the nucleosides of the antisense oligonucleotides are linked by phosphorothioate internucleoside linkages. In certain embodiments, the nucleosides of the antisense oligonucleotides are linked by phosphodiester internucleoside linkages. In certain embodiments, the nucleosides of the antisense oligonucleotides are linked by phosphorothioate and phosphodiester internucleotide linkages (e.g., the antisense oligonucleotides have “mixed backbones”).

In certain embodiments, nucleobases 135783-135929 are targeted by the following ISIS numbers: 424879, 424880, 548937, 613114-613119, 622096-622138, 623988-623996, 664511-664542, and 664661-664819.

In certain embodiments, nucleobases 135783-135929 are targeted by the following SEQ ID NOs: 56, 57, 248, 462-466, 1668-1686, 2025-2048, 2301-2309, 2331-2443, and 2478-2483.

In certain embodiments, antisense oligonucleotides targeting nucleobases 135783-135929 achieve at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31%, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41%, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, or at least 93% reduction of Tau mRNA and/or protein levels in vitro and/or in vivo.

4. Nucleobases 135783-135914 of SEQ ID NO: 1

In certain embodiments, antisense oligonucleotides are designed to target nucleobases 135783-135914 of SEQ ID NO: 1 (GENBANK Accession No. GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to 9381000). In certain embodiments, nucleobases 135783-135914 are a hotspot region. In certain embodiments, nucleobases 135783-135914 are targeted by antisense oligonucleotides. In certain embodiments, the antisense oligonucleotides are 18, 19, or 20 nucleobases in length. In certain embodiments, the antisense oligonucleotides are gapmers. In certain embodiments, the gapmers are 5-10-5 MOE gapmers, 5-9-5 MOE gapmers, 5-7-6 MOE gapmers, or 5-8-5 MOE gapmers. In certain embodiments, the nucleosides of the antisense oligonucleotides are linked by phosphorothioate internucleoside linkages. In certain embodiments, the nucleosides of the antisense oligonucleotides are linked by phosphodiester internucleoside linkages. In certain embodiments, the nucleosides of the antisense oligonucleotides are linked by phosphorothioate and phosphodiester internucleotide linkages (e.g., the antisense oligonucleotides have “mixed backbones”).

In certain embodiments, nucleobases 135783-135914 are targeted by the following ISIS numbers: 424879, 424880, 548937, 613114-613119, 622096-622133, 623988-623996, 664511-664542, 664661-664819.

In certain embodiments, nucleobases 135783-135914 are targeted by the following SEQ ID NOs: 56, 57, 248, 462-466, 1668-1681, 2025-2048, 2301-2309, 2331-2443, and 2478-2483.

In certain embodiments, nucleobases 135783-135914 are targeted by the following ISIS numbers: 424879, 424880, 548937, 613114-613119, 622096-622133, and 623988-623996.

In certain embodiments, nucleobases 135783-135914 are targeted by the following SEQ ID NOs: 56, 57, 248, 462-466, 1668-1681, 2025-2048, and 2301-2309.

In certain embodiments, antisense oligonucleotides targeting nucleobases 135783-135914 achieve at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31%, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41%, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, or at least 93% reduction of Tau mRNA and/or protein levels in vitro and/or in vivo.

EXAMPLES

Non-Limiting Disclosure and Incorporation by Reference

While certain compounds, compositions, and methods described herein have been described with specificity in accordance with certain embodiments, the following examples serve only to illustrate the compounds described herein and are not intended to limit the same. Each of the references recited in the present application is incorporated herein by reference in its entirety.

Example 1 Antisense Inhibition of Human Tau in HepG2 Cells by MOE Gapmers

Antisense oligonucleotides were designed targeting a tau nucleic acid and were tested for their effects on tau mRNA in vitro. Cultured HepG2 cells were transfected using Lipofectin reagent with 100 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and tau mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3104 (forward sequence AAGATTGGGTCCCTGGACAAT, designated herein as SEQ ID NO: 10; reverse sequence AGCTTGTGGGTTTCAATCTTTTTATT, designated herein as SEQ ID NO: 11; probe sequence CACCCACGTCCCTGGCGGA, designated herein as SEQ ID NO: 12) was used to measure mRNA levels. Tau mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of tau, relative to untreated control cells.

The newly designed chimeric antisense oligonucleotides in the Tables below were designed as 5-10-5 MOE gapmers. The gapmers are 20 nucleosides in length, wherein the central gap segment comprises ten 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five nucleosides each. Each nucleoside in the 5′ wing segment and each nucleoside in the 3′ wing segment has a 2′-MOE modification. The internucleoside linkages throughout each gapmer are phosphorothioate linkages. All cytosine residues throughout each gapmer are 5-methylcytosines. “Start site” indicates the 5′-most nucleoside to which the gapmer is targeted in the human gene sequence. “Stop site” indicates the 3′-most nucleoside to which the gapmer is targeted human gene sequence. Each gapmer listed in Table 1 below is targeted to either the human tau genomic sequence, designated herein as SEQ ID NO: 1 (GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to 9381000) or to the human tau mRNA sequence, designated herein as SEQ ID NO: 2 (GENBANK Accession No. NM_001123066.3). ‘n/a’ indicates that the oligonucleotide does not target the gene sequence with 100% complementarity. The sequences listed in Table 2 do not target either SEQ ID NO: 1 or 2 with 100% complementarity, but instead target SEQ ID NO: 3 (GENBANK Accession No. NM_016841.4, a variant mRNA sequence which skips exons 3, 4, 6, 8, 10, and 12) or SEQ ID NO: 4 (GENBANK Accession No. NT_010783.14 truncated from nucleotides 2624000 to 2761000).

TABLE 1 Inhibition of Tau mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2 SEQ SEQ SEQ SEQ ID ID ID ID NO: 1 NO: 1 NO: 2 NO: 2 SEQ ISIS Start Stop % Start Stop ID NO Site Site Sequence inhibition Site Site NO 424863 n/a n/a TTGTAGACTATTTGCACACT 59 2240 2259 20 433519 28769 28788 CACAGGCAGATGCGAACCCT 57 n/a n/a 21 433520 47181 47200 TGGTGGAGACAAGACATTCT 57 n/a n/a 22 433521 71387 71406 CCATCCCCTAATAGTTAGCA 29 n/a n/a 23 433522 72861 72880 CATGAGGCTTGGGATCTGAA 49 n/a n/a 24 433474 73879 73898 GTGATCTTCCATCACTTCGA 61 345 364 25 433523 73980 73999 GTCCACTAACCTTTCAGGCC 63 n/a n/a 26 433518 83423 83442 AGCATCAGAGGTTTCAGAGC 46  501  520 27 424852 83437 83456 GTTGGAGTGCTCTTAGCATC 58  515  534 28 433513 85934 85953 GGCAGCCTGCTTGCCGGGAG 68  573  592 29 433524 87927 87946 GAGGATTTCCTTGGAGAGAG 53 n/a n/a 30 433525 89950 89969 GTTCACTGACCTTGGGTCAC 38 n/a n/a 31 433526 91248 91267 ATGATTTCTAGAGGTCATGC 61 n/a n/a 32 433514 95121 95140 AGAGGAAATCCACAGGGAGG 43 1120 1139 33 433515 95372 95391 TTCAGAGGGCTCTGGAAGGT 58 1371 1390 34 433527 95790 95809 ACACCATGAGGGCACCCGTC 65 n/a n/a 35 433528 98549 98568 ACCATGCGAGCTGATAAAAT 46 n/a n/a 36 433516 101406 101425 AAGGTTTTAGCAGAGGAACG 50 1514 1533 37 433517 101507 101526 AGGAAGGTGGCTCTGGGCAC 73 1615 1634 38 433476 103087 103106 CCAGAGCTGGGTGGTGTCTT 60 n/a n/a 39 433477 108040 108059 GGTGGAGTACGGACCACTGC 71 2006 2025 40 433478 108054 108073 AAGACGGCGACTTGGGTGGA 65 2020 2039 41 424857 108148 108167 TGGTGCTTCAGGTTCTCAGT 54 2114 2133 42 433529 121819 121838 TTATCTGCACCTTTGGTAGC 29 n/a n/a 43 424859 121828 121847 TCTTATTAATTATCTGCACC 50 2149 2168 44 424860 121838 121857 AGATCCAGCTTCTTATTAAT 36 2159 2178 45 424861 121846 121865 CGTTGCTAAGATCCAGCTTC 48 2167 2186 46 424862 121865 121884 GAGCCACACTTGGACTGGAC 79 2186 2205 47 433530 122497 122516 GGTGGCGCAGGCTAAGCATA 52 n/a n/a 48 424864 125798 125817 GAGCCACACTTGGAGGTCAC 63 2279 2298 49 433531 125834 125853 ACAGGGCTACCTGGTTTATG 53 n/a n/a 50 424866 130141 130160 ATTTTACTTCCACCTGGCCA 70 2329 2348 51 433479 130188 130207 GGACCCAATCTTCGACTGGA 70 2376 2395 52 424926 135467 135486 GTGGGTTTCAATCTGCAAGA 39 n/a n/a 53 424869 135475 135494 GTCAGCTTGTGGGTTTCAAT 74 2438 2457 54 433480 135673 135692 GATCACAAACCCTGCTTGGC 60 2636 2655 55 424879 135848 135867 TGATTTTGAAGTCCCGAGCC 64 2811 2830 56 424880 135853 135872 ATCACTGATTTTGAAGTCCC 54 2816 2835 57 433481 136033 136052 CAGAAGCAGCTTTCAGAGCC 38 2996 3015 58 433482 136118 136137 AAATCCTTTGTTGCTGCCAC 37 3081 3100 59 424882 136425 136444 CCCACAGGCTGCCCTGCAGA 57 3388 3407 60 433483 136491 136510 GGAGGTCATCCACGAAGTGC 67 3454 3473 61 433484 136577 136596 GGAAGCCCCTCAACTCAGGC 28 3540 3559 62 433485 136655 136674 GGTCTGCAAAGTGGCCAAAA 49 3618 3637 63 424883 136675 136694 TGGTTAGCCCTAAAGTCCCA 54 3638 3657 64 433486 136686 136705 ACAAAGAGAACTGGTTAGCC 39 3649 3668 65 424884 136703 136722 AAGAGGCACAAGTCCTTACA 40 3666 3685 66 433487 136748 136767 CAGAGATGCCAGTGGCCCAG 70 3711 3730 67 433488 137083 137102 GGAACCGAATCAGATCATGA 54 4046 4065 68 433489 137387 137406 TGGAGGGCTGATACTATGCA 45 4350 4369 69 433490 137430 137449 AAAAGAACCATTTCCAAGGG 41 4393 4412 70 433491 137540 137559 ATCCAACTACAACTCAACAG 17 4503 4522 71 424894 137579 137598 ATCATAGTCACTCTGGTGAA 56 4542 4561 72 433492 137694 137713 GCCACACGAGTCCCAGTGTG 45 4657 4676 73 433493 137731 137750 AAAACTTGGGAGGCCCCAGC 50 4694 4713 74 433494 138173 138192 GATCCAGGACAGGCAATTCA 58 5136 5155 75 433495 138205 138224 TCCTCAGGCAGGCAGCTTGG 49 5168 5187 76 433496 138303 138322 TTCTCATGGCAGCAGATGGA 51 5266 5285 77 433497 138338 138357 TTAGGCAGCAATGTTTTGCA 18 5301 5320 78 433498 138503 138522 AGAGTTCTGGGCCCAGAGAC 73 5466 5485 79 433499 138675 138694 AGGAAGAGGAACCGAGGTGC 59 5638 5657 80 433500 138774 138793 TCTTAGGCTGGCCCCAAGAG 36 5737 5756 81 433501 138812 138831 TCAATTTATCTGCCAGCACT 28 5775 5794 82 433502 138845 138864 TCCTCATTTAAGATCACAAG 36 5808 5827 83 433503 138983 139002 ATGGAACTATTGATAAAGTG 47 5946 5965 84 433504 139003 139022 CACCACTGAAGTCAATTTAA 23 5966 5985 85 433505 139112 139131 AGTTTAAGTGCTGCACCCCA 52 6075 6094 86 433506 139137 139156 GAAATCATGAAAAGGGTTAC 25 6100 6119 87 433507 139154 139173 CTCTAGCAAATGTGGTTGAA 42 6117 6136 88 433508 139229 139248 AGCCAGCTGCCTGGGAAAGC 70 6192 6211 89 433509 139359 139378 TAGAGGGAAGGATGCCAAGG 56 6322 6341 90 433510 139406 139425 GTGTGTCTGGAGCCAGTGTG 49 6369 6388 91 424910 139675 139694 GAAATCATGGGACTTGCAAG 52 6638 6657 92 433512 139729 139748 AAAGCTAAGCTAAGATGATT 31 6692 6711 93 424913 139797 139816 TTACAGCAACAGTCAGTGTA 42 6760 6779 94

TABLE 2 Inhibition of Tau mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 3 and 4 Target SEQ Target SEQ ISIS ID Start % ID NO NO Site Sequence inhibition NO 424917 3 968 TTGTAGACTATTTGCA 51 95 CCTT 433475 3 443 TCTTCAGCTTTCAGGC 68 96 CAGC 433511 4 135519 AGAAGTTTTATGAAGC 13 97 CGCA

Example 2 Dose-Dependent Antisense Inhibition of Human Tau in HepG2 Cells by 5-10-5 MOE Gapmers

Gapmers from studies described above exhibiting significant in vitro inhibition of Tau mRNA were selected and tested at various doses in HepG2 cells. Cells were plated at a density of 10,000 cells per well and transfected using Lipofectin reagent with 12.5 nM, 25.0 nM, 50.0 nM, 100.0 nM, or 200.0 nM concentrations of antisense oligonucleotide, as specified in the Table below. After a treatment period of approximately 16 hours, RNA was isolated from the cells and Tau mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3104 was used to measure mRNA levels. Tau mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of tau, relative to untreated control cells. Tau mRNA levels were significantly reduced in a dose-dependent manner in antisense oligonucleotide treated cells.

TABLE 3 ISIS No 12.5 nM 25.0 nM 50.0 nM 100.0 nM 200.0 nM 424862 17 24 41 64 84 424866 14 25 47 63 84 433498 27 42 61 79 82 433508 25 31 53 66 83 433517 25 34 57 72 78 433487 18 30 38 60 76 433477 23 31 51 71 80 433475 18 33 57 71 85 433513 24 30 50 71 77 433483 23 28 41 58 74 433527 19 32 46 60 77

Example 3 Antisense Inhibition of Human Tau in SH-SY5Y Cells by 5-10-5 MOE Gapmers

Additional antisense oligonucleotides were designed targeting a Tau nucleic acid and were tested for their effects on Tau mRNA in vitro. The antisense oligonucleotides were tested in a series of experiments that had similar culture conditions. The results for each experiment are presented in separate tables shown below. Cultured SH-SY5Y cells were transfected using electroporation with 7,000 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and Tau mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3104 was used to measure mRNA levels. Tau mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of Tau, relative to untreated control cells.

The newly designed chimeric antisense oligonucleotides in the Tables below were designed as 5-10-5 MOE gapmers. The gapmers are 20 nucleosides in length, wherein the central gap segment comprises ten 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five nucleosides each. Each nucleoside in the 5′ wing segment and each nucleoside in the 3′ wing segment has a 2′-MOE modification. The internucleoside linkages throughout each gapmer are phosphorothioate linkages. All cytosine residues throughout each gapmer are 5-methylcytosines. “Start site” indicates the 5′-most nucleoside to which the gapmer is targeted in the human gene sequence. “Stop site” indicates the 3′-most nucleoside to which the gapmer is targeted human gene sequence. Each gapmer listed in the Tables below is targeted to either the human Tau genomic sequence, designated herein as SEQ ID NO: 1 (GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to 9381000) or to the human Tau mRNA sequence, designated herein as SEQ ID NO: 2 (GENBANK Accession No. NM_001123066.3). ‘n/a’ indicates that the antisense oligonucleotide does not target that particular gene sequence with 100% complementarity.

TABLE 4 SEQ SEQ SEQ SEQ ID ID ID ID NO: 1 NO: 1 NO: 2 NO: 2 SEQ ISIS Start Stop % Start Stop ID NO Site Site Sequence inhibition Site Site NO 548786 73867 73886 CACTTCGAACTCCTGGCGGG 73 333 352 98 548787 73869 73888 ATCACTTCGAACTCCTGGCG 64 335 354 99 548788 73871 73890 CCATCACTTCGAACTCCTGG 78 337 356 100 548789 73873 73892 TTCCATCACTTCGAACTCCT 49 339 358 101 548790 73875 73894 TCTTCCATCACTTCGAACTC 33 341 360 102 548791 73877 73896 GATCTTCCATCACTTCGAAC 54 343 362 103 433474 73879 73898 GTGATCTTCCATCACTTCGA 86 345 364 25 548792 73906 73925 CCTGTCCCCCAACCCGTACG 38 372 391 104 548793 73908 73927 TTCCTGTCCCCCAACCCGTA 58 374 393 105 548794 73910 73929 CTTTCCTGTCCCCCAACCCG 55 376 395 106 548795 73912 73931 ATCTTTCCTGTCCCCCAACC 14 378 397 107 548796 73914 73933 TGATCTTTCCTGTCCCCCAA 59 380 399 108 548797 73916 73935 CCTGATCTTTCCTGTCCCCC 67 382 401 109 548798 73918 73937 CCCCTGATCTTTCCTGTCCC 67 384 403 110 548799 73920 73939 CCCCCCTGATCTTTCCTGTC 33 386 405 111 548800 73922 73941 AGCCCCCCTGATCTTTCCTG 45 388 407 112 548801 73924 73943 GTAGCCCCCCTGATCTTTCC 62 390 409 113 548802 73926 73945 GTGTAGCCCCCCTGATCTTT 42 392 411 114 548803 73928 73947 TGGTGTAGCCCCCCTGATCT 58 394 413 115 548804 73930 73949 CATGGTGTAGCCCCCCTGAT 62 396 415 116 548805 73932 73951 TGCATGGTGTAGCCCCCCTG 82 398 417 117 548806 73934 73953 GGTGCATGGTGTAGCCCCCC 79 400 419 118 548807 73936 73955 TTGGTGCATGGTGTAGCCCC 58 402 421 119 548808 73938 73957 TCTTGGTGCATGGTGTAGCC 60 404 423 120 548809 73940 73959 GGTCTTGGTGCATGGTGTAG 67 406 425 121 548810 73945 73964 CTCTTGGTCTTGGTGCATGG 61 411 430 122 548811 73958 73977 CGTCCGTGTCACCCTCTTGG 73 424 443 123 548812 98559 98578 GCTTTTACTGACCATGCGAG 68 1449 1468 124 548813 98561 98580 TTGCTTTTACTGACCATGCG 64 1451 1470 125 548814 98563 98582 CTTTGCTTTTACTGACCATG 40 1453 1472 126 548815 98565 98584 GTCTTTGCTTTTACTGACCA 60 1455 1474 127 548816 98567 98586 CCGTCTTTGCTTTTACTGAC 50 1457 1476 128 548817 98569 98588 TCCCGTCTTTGCTTTTACTG 46 1459 1478 129 548818 98571 98590 AGTCCCGTCTTTGCTTTTAC 41 1461 1480 130 548819 98573 98592 CCAGTCCCGTCTTTGCTTTT 37 1463 1482 131 548820 98577 98596 GCTTCCAGTCCCGTCTTTGC 43 1467 1486 132 548821 98579 98598 TCGCTTCCAGTCCCGTCTTT 35 1469 1488 133 548822 98581 98600 CATCGCTTCCAGTCCCGTCT 73 1471 1490 134 548823 98583 98602 GTCATCGCTTCCAGTCCCGT 63 1473 1492 135 548824 98585 98604 TTGTCATCGCTTCCAGTCCC 50 1475 1494 136 548825 98587 98606 TTTTGTCATCGCTTCCAGTC 46 1477 1496 137 548826 103023 103042 GGCCCTTCTGGCCTGGAGGG 64 1747 1766 138 548827 103025 103044 CTGGCCCTTCTGGCCTGGAG 29 1749 1768 139 548828 103027 103046 GCCTGGCCCTTCTGGCCTGG 40 1751 1770 140 548829 103029 103048 TGGCCTGGCCCTTCTGGCCT 12 1753 1772 141 548830 103031 103050 GTTGGCCTGGCCCTTCTGGC 53 1755 1774 142 548831 103052 103071 TTTTGCTGGAATCCTGGTGG 20 1776 1795 143 548832 103054 103073 GTTTTTGCTGGAATCCTGGT 29 1778 1797 144 548833 103056 103075 GGGTTTTTGCTGGAATCCTG 51 1780 1799 145 548834 103075 103094 GGTGTCTTTGGAGCGGGCGG 44 1799 1818 146 548835 103077 103096 GTGGTGTCTTTGGAGCGGGC 30 1801 1820 147 548836 103079 103098 GGGTGGTGTCTTTGGAGCGG 38 1803 1822 148 548837 103081 103100 CTGGGTGGTGTCTTTGGAGC 29 1805 1824 149 548838 103083 103102 AGCTGGGTGGTGTCTTTGGA 37 1807 1826 150 548839 103085 103104 AGAGCTGGGTGGTGTCTTTG 0 1809 1828 151 548840 107921 107940 ATCCCCTGATTTTGGAGGTT 27 1887 1906 152 548841 107923 107942 CGATCCCCTGATTTTGGAGG 65 1889 1908 153 548842 107925 107944 TGCGATCCCCTGATTTTGGA 53 1891 1910 154 548843 107927 107946 GCTGCGATCCCCTGATTTTG 23 1893 1912 155 548844 107929 107948 CCGCTGCGATCCCCTGATTT 32 1895 1914 156 548845 107931 107950 AGCCGCTGCGATCCCCTGAT 51 1897 1916 157 548846 107933 107952 GTAGCCGCTGCGATCCCCTG 14 1899 1918 158 548847 107968 107987 CGGCTGCCGGGAGTGCCTGG 33 1934 1953 159 548848 107970 107989 AGCGGCTGCCGGGAGTGCCT 52 1936 1955 160 548849 107972 107991 GGAGCGGCTGCCGGGAGTGC 44 1938 1957 161 548850 108047 108066 CGACTTGGGTGGAGTACGGA 10 2013 2032 162 548851 108049 108068 GGCGACTTGGGTGGAGTACG 29 2015 2034 163 548852 108051 108070 ACGGCGACTTGGGTGGAGTA 39 2017 2036 164 548853 108053 108072 AGACGGCGACTTGGGTGGAG 6 2019 2038 165 548854 108055 108074 GAAGACGGCGACTTGGGTGG 0 2021 2040 166 548855 108057 108076 CGGAAGACGGCGACTTGGGT 40 2023 2042 167 548856 108059 108078 GGCGGAAGACGGCGACTTGG 38 2025 2044 168 548857 108061 108080 TTGGCGGAAGACGGCGACTT 41 2027 2046 169 548858 108063 108082 TCTTGGCGGAAGACGGCGAC 35 2029 2048 170 548859 108065 108084 GCTCTTGGCGGAAGACGGCG 54 2031 2050 171 548860 108067 108086 CGGCTCTTGGCGGAAGACGG 58 2033 2052 172 548861 108069 108088 GGCGGCTCTTGGCGGAAGAC 55 2035 2054 173 433498 138503 138522 AGAGTTCTGGGCCCAGAGAC 53 5466 5485 79

TABLE 5 SEQ SEQ SEQ SEQ ID ID ID ID NO: 1 NO: 1 NO: 2 NO: 2 ISIS Start Stop % Start Stop SEQ NO Site Site Sequence inhibition Site Site ID NO 548885 n/a n/a GGGAGATTCTTTCAGGCCAG 0 444 463 174 548893 n/a n/a GAGCTTTGAGTTGAGGGACC 0 1432 1451 175 548894 n/a n/a GACCATGCGAGCTTTGAGTT 13 1440 1459 176 548895 n/a n/a TACTGACCATGCGAGCTTTG 33 1444 1463 177 548881 6183 6202 ATAGTCGACAGAGGCGAGGA 43 283 302 178 548882 73840 73859 CATCCTGGTTCAAAGTTCAC 30 306 325 179 548883 73846 73865 CTCAGCCATCCTGGTTCAAA 39 312 331 180 548884 73847 73866 GCTCAGCCATCCTGGTTCAA 53 313 332 181 548886 83401 83420 GGTTCCTCAGATCCGTCCTC 53 479 498 182 548887 83438 83457 TGTTGGAGTGCTCTTAGCAT 33 516 535 183 548888 85939 85958 TGCGCGGCAGCCTGCTTGCC 32 578 597 184 548889 94699 94718 ACCACCTTACCACTTTCAGG 35 698 717 185 548890 94775 94794 GGGAGCCCCAGGCATGCCGG 45 774 793 186 548891 95125 95144 TTGGAGAGGAAATCCACAGG 13 1124 1143 187 548892 95131 95150 GAAACTTTGGAGAGGAAATC 0 1130 1149 188 548896 98557 98576 TTTTACTGACCATGCGAGCT 40 1447 1466 189 548897 101425 101444 AAGGCAAGGCCTATTTTTCA 34 1533 1552 190 548898 101469 101488 TGGATCAGAGGGTCTGAGCT 40 1577 1596 191 548899 101501 101520 GTGGCTCTGGGCACACAGCA 63 1609 1628 192 548900 108037 108056 GGAGTACGGACCACTGCCAC 37 2003 2022 193 433477 108040 108059 GGTGGAGTACGGACCACTGC 21 2006 2025 40 548862 108071 108090 CAGGCGGCTCTTGGCGGAAG 24 2037 2056 194 548863 108073 108092 TGCAGGCGGCTCTTGGCGGA 34 2039 2058 195 548864 108075 108094 TCTGCAGGCGGCTCTTGGCG 40 2041 2060 196 548865 108129 108148 TGGAGCCGATCTTGGACTTG 23 2095 2114 197 548866 108131 108150 AGTGGAGCCGATCTTGGACT 4 2097 2116 198 548901 108134 108153 CTCAGTGGAGCCGATCTTGG 28 2100 2119 199 548902 108136 108155 TTCTCAGTGGAGCCGATCTT 9 2102 2121 200 548903 108155 108174 TCCCGGCTGGTGCTTCAGGT 33 2121 2140 201 548904 121675 121694 TTTCGGACTTGGCAGAGGCA 50 n/a n/a 202 548905 121728 121747 GAGTGACATGCGCCACCCTG 65 n/a n/a 203 548906 121736 121755 CTTTCGATGAGTGACATGCG 19 n/a n/a 204 548907 121794 121813 AAAAGGATGAGTGACACGCC 58 n/a n/a 205 548908 121916 121935 GTGAAGGTACTCACACTGCC 34 n/a n/a 206 548909 121917 121936 TGTGAAGGTACTCACACTGC 21 n/a n/a 207 548910 121992 122011 CTATGCAGTGTCTCGCAAGT 39 n/a n/a 208 548911 122001 122020 GGATTTATTCTATGCAGTGT 38 n/a n/a 209 548912 122002 122021 AGGATTTATTCTATGCAGTG 37 n/a n/a 210 548913 122015 122034 TGAGAGCCCAAGAAGGATTT 15 n/a n/a 211 548914 122022 122041 CAGATCCTGAGAGCCCAAGA 33 n/a n/a 212 548915 122025 122044 AGCCAGATCCTGAGAGCCCA 55 n/a n/a 213 548916 122137 122156 GGACCCAGAAATGCTGGGAC 18 n/a n/a 214 548917 125555 125574 AAATGTGTTGTCGAAATTCT 0 n/a n/a 215 548918 125568 125587 AGTCAGGGTGGAAAAATGTG 0 n/a n/a 216 548919 125578 125597 AGCACATCCTAGTCAGGGTG 39 n/a n/a 217 548920 125587 125606 GCCATGAGGAGCACATCCTA 45 n/a n/a 218 548921 125600 125619 GTGGTTCCCAGCTGCCATGA 38 n/a n/a 219 548922 125603 125622 ACAGTGGTTCCCAGCTGCCA 38 n/a n/a 220 548923 125619 125638 CCCAGGCCCTTATTGGACAG 19 n/a n/a 221 548867 125778 125797 CTTGCTCAGGTCAACTGGTT 45 2259 2278 222 548925 125812 125831 GGATGTTGCCTAATGAGCCA 35 2293 2312 223 548926 130194 130213 GTCCAGGGACCCAATCTTCG 87 2382 2401 224 548927 130196 130215 TTGTCCAGGGACCCAATCTT 48 2384 2403 225 548928 135478 135497 AAGGTCAGCTTGTGGGTTTC 52 2441 2460 226 548868 135518 135537 CTCCGCCCCGTGGTCTGTCT 37 2481 2500 227 548869 135520 135539 ATCTCCGCCCCGTGGTCTGT 25 2483 2502 228 548870 135522 135541 CGATCTCCGCCCCGTGGTCT 48 2485 2504 229 548871 135524 135543 CACGATCTCCGCCCCGTGGT 38 2487 2506 230 548872 135526 135545 TACACGATCTCCGCCCCGTG 33 2489 2508 231 548873 135528 135547 TGTACACGATCTCCGCCCCG 22 2491 2510 232 548874 135530 135549 CTTGTACACGATCTCCGCCC 38 2493 2512 233 548875 135532 135551 GACTTGTACACGATCTCCGC 13 2495 2514 234 548876 135534 135553 GCGACTTGTACACGATCTCC 40 2497 2516 235 548877 135536 135555 TGGCGACTTGTACACGATCT 33 2499 2518 236 548878 135538 135557 ACTGGCGACTTGTACACGAT 25 2501 2520 237 548879 135540 135559 CCACTGGCGACTTGTACACG 19 2503 2522 238 548929 135546 135565 CAGACACCACTGGCGACTTG 33 2509 2528 239 548930 135593 135612 GCTGCCGGTGGAGGAGACAT 29 2556 2575 240 548931 135594 135613 TGCTGCCGGTGGAGGAGACA 19 2557 2576 241 548932 135600 135619 TGTCGATGCTGCCGGTGGAG 20 2563 2582 242 548933 135601 135620 ATGTCGATGCTGCCGGTGGA 20 2564 2583 243 548934 135610 135629 GAGTCTACCATGTCGATGCT 23 2573 2592 244 548880 135654 135673 CCAGGGAGGCAGACACCTCG 28 2617 2636 245 548935 135662 135681 CTGCTTGGCCAGGGAGGCAG 14 2625 2644 246 548936 135674 135693 TGATCACAAACCCTGCTTGG 25 2637 2656 247 548937 135786 135805 CCGAACTGCGAGGAGCAGCT 66 2749 2768 248 433498 138503 138522 AGAGTTCTGGGCCCAGAGAC 34 5466 5485 79

Example 4 Dose-Dependent Antisense Inhibition of Human Tau in SH-SY5Y Cells by 5-10-5 MOE Gapmers

Gapmers from studies described above exhibiting significant in vitro inhibition of Tau mRNA were selected and tested at various doses in SH-SY-5Y cells. Cells were plated at a density of 20,000 cells per well and transfected using electroporation with 1.25 μM, 2.50 μM, 5.00 μM, 10.00 μM, and 20.00 μM concentrations of antisense oligonucleotide, as specified in the Table below. After a treatment period of approximately 16 hours, RNA was isolated from the cells and Tau mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3104 was used to measure mRNA levels. Tau mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of Tau, relative to untreated control cells. Tau mRNA levels were significantly reduced in a dose-dependent manner in antisense oligonucleotide treated cells.

TABLE 6 ISIS No 1.25 μM 2.50 μM 5.00 μM 10.00 μM 20.00 μM 433474 29 54 74 83 88 433477 0 0 11 33 59 433498 0 10 30 53 71 548786 9 29 51 66 82 548787 19 27 61 85 87 548788 31 31 54 72 94 548805 19 48 62 81 96 548806 25 50 75 80 95 548811 21 42 60 74 94 548812 47 39 64 77 91 548822 0 32 52 62 91 548899 19 50 69 87 93 548905 11 32 57 80 95 548907 14 28 44 80 91 548937 24 43 66 82 91

Example 5 Antisense Inhibition of Human Tau in SH-SY5Y Cells by 5-10-5 MOE Gapmers

Additional antisense oligonucleotides were designed targeting a Tau nucleic acid and were tested for their effects on Tau mRNA in vitro. Cultured SH-SY5Y cells were plated at a density of 20,000 cells per well and were transfected using electroporation with 6,000 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and Tau mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3104 was used to measure mRNA levels. Tau mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of tau, relative to untreated control cells.

The newly designed chimeric antisense oligonucleotides in the Tables below were designed as 5-10-5 MOE gapmers. The gapmers are 20 nucleosides in length, wherein the central gap segment comprises of ten 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five nucleosides each. Each nucleoside in the 5′ wing segment and each nucleoside in the 3′ wing segment has a 2′-MOE modification. The internucleoside linkages throughout each gapmer are phosphorothioate linkages. All cytosine residues throughout each gapmer are 5-methylcytosines. “Start site” indicates the 5′-most nucleoside to which the gapmer is targeted in the human gene sequence. “Stop site” indicates the 3′-most nucleoside to which the gapmer is targeted human gene sequence. Each gapmer listed in the Tables below is targeted to the human tau genomic sequence, designated herein as SEQ ID NO: 1 (GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to 9381000).

TABLE 7 SEQ ID SEQ ID ISIS NO: 1 NO: 1 % SEQ NO Start Site Stop Site Sequence inhibition ID NO 433474 73879 73898 GTGATCTTCCATCACTTCGA 63 25 559166 26760 26779 TGCTTACACACCACACACAC 0 249 26944 26963 559167 26761 26780 ATGCTTACACACCACACACA 4 250 26945 26964 559168 26762 26781 CATGCTTACACACCACACAC 0 251 26946 26965 559169 81022 81041 TACAGATATATGATTGGATG 0 252 81125 81144 559170 81023 81042 GTACAGATATATGATTGGAT 0 253 81126 81145 559171 81024 81043 TGTACAGATATATGATTGGA 0 254 81127 81146 559172 81025 81044 ATGTACAGATATATGATTGG 0 255 81128 81147 559173 81026 81045 TATGTACAGATATATGATTG 0 256 81129 81148 559174 81027 81046 TTATGTACAGATATATGATT 0 257 81130 81149 559175 81028 81047 ATTATGTACAGATATATGAT 0 258 81131 81150 559176 81029 81048 GATTATGTACAGATATATGA 0 259 81132 81151 559177 81030 81049 GGATTATGTACAGATATATG 0 260 81133 81152 559178 81031 81050 TGGATTATGTACAGATATAT 0 261 81134 81153 559179 81032 81051 ATGGATTATGTACAGATATA 0 262 81135 81154 559180 81033 81052 AATGGATTATGTACAGATAT 0 263 81136 81155 559181 81034 81053 GAATGGATTATGTACAGATA 0 264 81137 81156 559182 81035 81054 AGAATGGATTATGTACAGAT 0 265 81138 81157 559183 81036 81055 AAGAATGGATTATGTACAGA 0 266 81139 81158 559184 81037 81056 GAAGAATGGATTATGTACAG 0 267 81140 81159 559185 81038 81057 GGAAGAATGGATTATGTACA 0 268 81141 81160 559186 81039 81058 GGGAAGAATGGATTATGTAC 0 269 81142 81161 559187 81040 81059 AGGGAAGAATGGATTATGTA 0 270 81143 81162 559188 81041 81060 GAGGGAAGAATGGATTATGT 0 271 81144 81163 559189 81042 81061 CGAGGGAAGAATGGATTATG 0 272 81145 81164 559190 81043 81062 CCGAGGGAAGAATGGATTAT 33 273 81146 81165 559191 81044 81063 ACCGAGGGAAGAATGGATTA 12 274 81147 81166 559192 81045 81064 AACCGAGGGAAGAATGGATT 1 275 81148 81167 559193 81046 81065 GAACCGAGGGAAGAATGGAT 0 276 81149 81168 559194 81047 81066 TGAACCGAGGGAAGAATGGA 0 277 81150 81169 559195 81048 81067 ATGAACCGAGGGAAGAATGG 0 278 81151 81170 559196 81049 81068 GATGAACCGAGGGAAGAATG 0 279 81152 81171 559197 81050 81069 GGATGAACCGAGGGAAGAAT 0 280 81153 81172 559198 81051 81070 TGGATGAACCGAGGGAAGAA 0 281 81154 81173 559199 81052 81071 ATGGATGAACCGAGGGAAGA 0 282 81155 81174 559200 81053 81072 GATGGATGAACCGAGGGAAG 0 283 81156 81175 559201 81054 81073 GGATGGATGAACCGAGGGAA 1 284 81157 81176 559202 81055 81074 TGGATGGATGAACCGAGGGA 0 285 81158 81177 559203 81056 81075 ATGGATGGATGAACCGAGGG 0 286 81159 81178 559204 81057 81076 GATGGATGGATGAACCGAGG 0 287 81160 81179 559205 81058 81077 GGATGGATGGATGAACCGAG 0 288 81161 81180 559206 81096 81115 AAGGATGAAGGAAGGGATGG 0 289 81199 81218 559207 81097 81116 GAAGGATGAAGGAAGGGATG 0 290 81200 81219 559208 81098 81117 GGAAGGATGAAGGAAGGGAT 0 291 81201 81220 559209 81100 81119 TAGGAAGGATGAAGGAAGGG 0 292 81203 81222 559210 81101 81120 ATAGGAAGGATGAAGGAAGG 0 293 81204 81223 559211 81102 81121 GATAGGAAGGATGAAGGAAG 0 294 81205 81224 559212 81103 81122 TGATAGGAAGGATGAAGGAA 0 295 81206 81225 559213 81104 81123 ATGATAGGAAGGATGAAGGA 0 296 81207 81226 559214 81105 81124 GATGATAGGAAGGATGAAGG 0 297 81208 81227 559215 81106 81125 GGATGATAGGAAGGATGAAG 0 298 81209 81228 559216 81107 81126 TGGATGATAGGAAGGATGAA 0 299 81210 81229 559217 81108 81127 ATGGATGATAGGAAGGATGA 0 300 81211 81230 559218 81109 81128 GATGGATGATAGGAAGGATG 0 301 81212 81231 559219 81110 81129 GGATGGATGATAGGAAGGAT 0 302 81213 81232 559220 81111 81130 TGGATGGATGATAGGAAGGA 0 303 81214 81233 559221 81112 81131 TTGGATGGATGATAGGAAGG 0 304 81215 81234 559222 81113 81132 ATTGGATGGATGATAGGAAG 0 305 81216 81235 559223 81114 81133 GATTGGATGGATGATAGGAA 0 306 81217 81236 559224 81115 81134 TGATTGGATGGATGATAGGA 0 307 81218 81237 559225 81116 81135 ATGATTGGATGGATGATAGG 0 308 81219 81238 559226 81117 81136 TATGATTGGATGGATGATAG 0 309 81220 81239 559227 80890 80909 AGATATGGGTGGATGGATGG 0 310 81472 81491 559228 80891 80910 AAGATATGGGTGGATGGATG 0 311 81473 81492 559229 80892 80911 GAAGATATGGGTGGATGGAT 0 312 81474 81493 559230 98881 98900 ACACACCTTCATTTACTGTC 79 313 98918 98937 559231 98882 98901 AACACACCTTCATTTACTGT 56 314 98919 98938 559232 98883 98902 AAACACACCTTCATTTACTG 59 315 98920 98939 559233 98884 98903 CAAACACACCTTCATTTACT 51 316 98921 98940 559234 98885 98904 TCAAACACACCTTCATTTAC 25 317 98922 98941 559235 98887 98906 TTTCAAACACACCTTCATTT 25 318 98924 98943 559236 98888 98907 TTTTCAAACACACCTTCATT 14 319 98925 98944 559237 98889 98908 GTTTTCAAACACACCTTCAT 42 320 98926 98945 559238 98890 98909 GGTTTTCAAACACACCTTCA 79 321 98927 98946 559239 98891 98910 TGGTTTTCAAACACACCTTC 87 322 98928 98947 559240 111795 111814 GAAAACAGAGAAAAGCACAA 12 323 111819 111838 559241 119594 119613 AGAAGTGATATCATATCCTA 63 324 119905 119924 559242 119595 119614 AAGAAGTGATATCATATCCT 74 325 119906 119925

Example 6 Dose-Dependent Antisense Inhibition of Human Tau in SH-SY5Y Cells by 5-10-5 MOE Gapmers

Gapmers from studies described above exhibiting significant in vitro inhibition of Tau mRNA were selected and tested at various doses in SH-SY-5Y cells. Cells were plated at a density of 20,000 cells per well and transfected using electroporation with 0.625 μM, 1.25 μM, 2.500 μM, 5.00 μM, 10.00 μM, and 20.00 μM concentrations of antisense oligonucleotide, as specified in the Table below. After a treatment period of approximately 16 hours, RNA was isolated from the cells and Tau mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3104 was used to measure mRNA levels. Tau mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of Tau, relative to untreated control cells. Tau mRNA levels were significantly reduced in a dose-dependent manner in antisense oligonucleotide treated cells.

TABLE 8 0.625 1.25 2.50 5.00 10.00 20.00 ISIS No μM μM μM μM μM μM 433474 17 39 53 73 81 88 559230 30 50 67 81 92 94 559231 0 16 26 44 63 83 559232 10 12 23 30 62 79 559233 5 10 21 38 62 81 559237 0 16 27 31 58 88 559238 25 18 72 77 86 86 559239 26 47 64 80 88 93 559241 0 18 34 61 74 95 559242 13 20 23 47 71 83

Example 7 Antisense Inhibition of Human Tau in SH-SY5Y Cells by MOE Gapmers with Phosphorothioate and Phosphodiester Internucleoside Linkages

Antisense oligonucleotides were designed targeting a tau nucleic acid and were tested for their effects on tau mRNA in vitro. The antisense oligonucleotides were tested in a series of experiments that had similar culture conditions. The results for each experiment are presented in separate tables shown below. Cultured SH-SY5Y cells were transfected using electroporation with 8,000 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and tau mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3104 was used to measure mRNA levels. Tau mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of tau, relative to untreated control cells.

The newly designed chimeric antisense oligonucleotides in the Tables below were designed as 5-10-5 MOE gapmers. The gapmers are 20 nucleosides in length, wherein the central gap segment comprises of ten 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five nucleosides each. Each nucleoside in the 5′ wing segment and each nucleoside in the 3′ wing segment has a 2′-MOE modification. The internucleoside linkages throughout each gapmer are either phosphorothioate linkages or phosphodiester linkages. All cytosine residues throughout each gapmer are 5-methylcytosines. The ‘Chemistry’ column describes the internucleoside linkages of each oligonucleotide. ‘s’ indicates phosphorothioate linkage and ‘o’ indicates phosphodiester linkage. “Start site” indicates the 5′-most nucleoside to which the gapmer is targeted in the human gene sequence. “Stop site” indicates the 3′-most nucleoside to which the gapmer is targeted human gene sequence.

Each gapmer listed in the Tables below is targeted to either the human Tau genomic sequence, designated herein as SEQ ID NO: 1 (GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to 9381000), human Tau mRNA sequences, designated herein as SEQ ID NO: 2 (GENBANK Accession No. NM_001123066.3) or SEQ ID NO: 3 (GENBANK Accession No. NM_016841.4). Several oligonucleotides, presented in Tables 10, 12, and 16, target variant mRNA sequences, designated herein as SEQ ID NO: 5 (GENBANK Accession No. DR002467.1), SEQ ID NO: 6 (GENBANK Accession No. NM_001203251.1) or SEQ ID NO: 7 (GENBANK Accession No. NM_016835.4). The oligonucleotides are presented in the various tables according to the main gene sequence that they target with 100% complementarity. ‘n/a’ indicates that the antisense oligonucleotide does not target that particular gene sequence with 100% complementarity.

TABLE 9 Inhibition of Tau mRNA by 5-10-5 MOE gapmers with phosphorothioate and phosphodiester internucleoside linkages targeting SEQ ID NOs: 1 and 3 SEQ SEQ SEQ SEQ ID ID ID ID NO: 1 NO: 1 NO: 3 NO: 3 SEQ ISIS Start Stop % Start Stop ID NO Site Site Sequence inhibition Site Site Linkage chemistry NO 433475 n/a n/a TCTTCAGCTTTCAGGCCAGC 41 443 462 sssssssssssssssssss 96 613242 n/a n/a CTTCAGCTTTCAGGCCAGCG 54 442 461 soooossssssssssooss 326 613243 n/a n/a TCTTCAGCTTTCAGGCCAGC 43 443 462 soooossssssssssooss 96 613244 n/a n/a CCATGCGAGCTTGGGTCACG 64 511 530 soooossssssssssooss 327 613245 n/a n/a CAGCCCCCTTGGCTTTTTTG 30 565 584 soooossssssssssooss 328 613247 n/a n/a TTGCACCTTCCCGCCTCCCG 53 957 976 soooossssssssssooss 329 613240 5897 5916 TGCCGCTCGGCCGTCCGGCG 9 n/a n/a soooossssssssssooss 330 613249 6292 6311 GCCCGGCGCACGAAGCCCCA 80 n/a n/a soooossssssssssooss 331 613250 7099 7118 AGAAAGAAATCCGCCCCGAG 45 n/a n/a soooossssssssssooss 332 613251 11709 11728 ATAATGGAATACTTATTGCA 54 n/a n/a soooossssssssssooss 333 613252 11819 11838 GCAGGAGCATGGCACCTGGA 79 n/a n/a soooossssssssssooss 334 613253 12081 12100 ACTCGCTGACATGGTGTTTG 87 n/a n/a soooossssssssssooss 335 613254 13386 13405 GATGACTCCACAATGTAAAC 75 n/a n/a soooossssssssssooss 336 613255 16760 16779 CCTATAGGATTATCCAGGAA 88 n/a n/a soooossssssssssooss 337 613256 16987 17006 CATTTCTCATCAGCCATCGA 81 n/a n/a soooossssssssssooss 338 613257 17428 17447 ACCAAAGACTTCAGATCAGC 88 n/a n/a soooossssssssssooss 339 613412 73879 73898 GTGATCTTCCATCACTTCGA 82 345 364 soooossssssssssooss 25 613246 103087 103106 CCAGAGCTGGGTGGTGTCTT 51 686 705 soooossssssssssooss 39 613181 138046 138065 AGCTTCTGGGAGCTGCAGAT 80 3737 3756 soooossssssssssooss 340 613182 138083 138102 AGAGGAACCCAGTCTGAGGG 49 3774 3793 soooossssssssssooss 341 613183 138116 138135 AGGCTGCGCTGCCCCTCCAG 68 3807 3826 soooossssssssssooss 342 613184 138149 138168 AATCCCTGCTGTGGTCGCAG 63 3840 3859 soooossssssssssooss 343 613185 138182 138201 TCTAGAGCAGATCCAGGACA 53 3873 3892 soooossssssssssooss 344 613186 138215 138234 AAGTCATCCTTCCTCAGGCA 63 3906 3925 soooossssssssssooss 345 613187 138248 138267 CAAGGCTTTGGGAACAGTGT 78 3939 3958 soooossssssssssooss 346 613188 138250 138269 GTCAAGGCTTTGGGAACAGT 84 3941 3960 soooossssssssssooss 347 613189 138281 138300 TTGTGCAAGGTCAGCGGGCT 75 3972 3991 soooossssssssssooss 348 613190 138314 138333 CGGCTTCCCTTTTCTCATGG 75 4005 4024 soooossssssssssooss 349 613191 138347 138366 TGAGTTTCTTTAGGCAGCAA 82 4038 4057 soooossssssssssooss 350 613192 138380 138399 AACCAGAAGTGGCAGAATTG 47 4071 4090 soooossssssssssooss 351 613193 138413 138432 CAAGTCCCTCAGGGTTGCCT 78 4104 4123 soooossssssssssooss 352 613194 138454 138473 CGAAGCTGCCAGCCCCAGGG 70 4145 4164 soooossssssssssooss 353 613195 138487 138506 AGACTTCCTTTCAGGTAAAG 77 4178 4197 soooossssssssssooss 354 613196 138503 138522 AGAGTTCTGGGCCCAGAGAC 58 4194 4213 soooossssssssssooss 79 613197 138520 138539 AGGGAGGCTCTTGGTGGAGA 35 4211 4230 soooossssssssssooss 355 613198 138553 138572 ACTTAGGAGAATTGCTGGGA 75 4244 4263 soooossssssssssooss 356 613199 138586 138605 CCCACATTTCCTTCTCCTTC 72 4277 4296 soooossssssssssooss 357 613200 138636 138655 ACTGTTGGCAGTAATGAGGG 71 4327 4346 soooossssssssssooss 358 613201 138674 138693 GGAAGAGGAACCGAGGTGCG 61 4365 4384 soooossssssssssooss 359 613202 138707 138726 CATGGTGCTGAAGAGCAGGG 71 4398 4417 soooossssssssssooss 360 613203 138740 138759 GGGAGATCCCAGAGCCTTCC 53 4431 4450 soooossssssssssooss 361 613204 138773 138792 CTTAGGCTGGCCCCAAGAGC 64 4464 4483 soooossssssssssooss 362 613205 138806 138825 TATCTGCCAGCACTGATCAC 56 4497 4516 soooossssssssssooss 363 613206 138839 138858 TTTAAGATCACAAGCCAGCG 62 4530 4549 soooossssssssssooss 364 613207 138872 138891 GAGGAGTGCCCAGCCCTGGG 54 4563 4582 soooossssssssssooss 365 613208 138905 138924 CTGGCTCTGCAGGTGGGAGA 44 4596 4615 soooossssssssssooss 366 613209 138938 138957 TACAGTATATCCTATCTAGC 57 4629 4648 soooossssssssssooss 367 613210 138971 138990 ATAAAGTGAGTCAGCAGCTT 71 4662 4681 soooossssssssssooss 368 613211 139004 139023 TCACCACTGAAGTCAATTTA 72 4695 4714 soooossssssssssooss 369 613212 139037 139056 AGCACAACAAGCAATAGCAA 80 4728 4747 soooossssssssssooss 370 613213 139070 139089 ACTATCTTACACATTCCTCC 48 4761 4780 soooossssssssssooss 371 613214 139103 139122 GCTGCACCCCAAGATCTCCC 79 4794 4813 soooossssssssssooss 372 613215 139136 139155 AAATCATGAAAAGGGTTACG 61 4827 4846 soooossssssssssooss 373 613216 139169 139188 GTGGCTGCTCCCTCCCTCTA 60 4860 4879 soooossssssssssooss 374 613217 139188 139207 CCCAAGGGCCTCTAACTCCG 64 4879 4898 soooossssssssssooss 375 613218 139202 139221 TGGAAAAGAGAAACCCCAAG 56 4893 4912 soooossssssssssooss 376 613219 139229 139248 AGCCAGCTGCCTGGGAAAGC 63 4920 4939 soooossssssssssooss 89 613220 139235 139254 TGAACTAGCCAGCTGCCTGG 46 4926 4945 soooossssssssssooss 377 613221 139268 139287 TATTCCTACGCCTGCACCTG 54 4959 4978 soooossssssssssooss 378 613222 139301 139320 AGAGGGCAGCAGGCCAAAGC 48 4992 5011 soooossssssssssooss 379 613223 139337 139356 TTAGGGAGGCATGATTGTGG 55 5028 5047 soooossssssssssooss 380 613224 139368 139387 CCAACGGCTTAGAGGGAAGG 61 5059 5078 soooossssssssssooss 381 613225 139370 139389 TGCCAACGGCTTAGAGGGAA 65 5061 5080 soooossssssssssooss 382 613226 139403 139422 TGTCTGGAGCCAGTGTGAGA 56 5094 5113 soooossssssssssooss 383 613227 139436 139455 CGAGTGATCTCAGCTCCAAA 79 5127 5146 soooossssssssssooss 384 613228 139502 139521 TGATCACCTCTGCCCTCGCC 72 5193 5212 soooossssssssssooss 385 613229 139535 139554 TGAAGCTGCAGGTCTGTAGA 57 5226 5245 soooossssssssssooss 386 613230 139568 139587 CTTTTCAAAGCTGAAGAGAA 57 5259 5278 soooossssssssssooss 387 613231 139601 139620 AGGTGAGGCTCTAGGCCAGT 76 5292 5311 soooossssssssssooss 388 613232 139636 139655 CAACATGGCAAACTCATGGG 76 5327 5346 soooossssssssssooss 389 613233 139669 139688 ATGGGACTTGCAAGTGCCAG 83 5360 5379 soooossssssssssooss 390 613234 139702 139721 CCTCCCCCCACCCTCAGAAT 15 5393 5412 soooossssssssssooss 391 613235 139735 139754 AGACAGAAAGCTAAGCTAAG 50 5426 5445 soooossssssssssooss 392 613236 139768 139787 TTAAAACACACAATACACTA 46 5459 5478 soooossssssssssooss 393 613237 139801 139820 ACTTTTACAGCAACAGTCAG 54 5492 5511 soooossssssssssooss 394 613238 139834 139853 TTTAATCAGAGTAATAACTT 12 5525 5544 soooossssssssssooss 395

TABLE 10 Inhibition of Tau mRNA by 5-10-5 MOE gapmers with phosphorothioate and phosphodiester internucleoside linkages targeting SEQ ID NOs: 5 and 6 Target SEQ Target SEQ ISIS ID Start % ID NO NO Site Sequence inhibition Linkage chemistry NO 613239 5 3 TGCCCTTCGCGGTCCCTTCG 2 soooossssssssssooss 396 613241 5 424 CTGTCCCCCAAACCCGTACG 56 soooossssssssssooss 397 613248 6 524 GCTTCCGCTGTTGGAGTGCT 62 soooossssssssssooss 398

TABLE 11 Inhibition of Tau mRNA by 5-10-5 MOE gapmers with phosphorothioate and phosphodiester internucleoside linkages targeting SEQ ID NOs: 1 and 3 SEQ SEQ SEQ SEQ ID ID ID ID NO: 1 NO: 1 NO: 3 NO: 3 SEQ ISIS Start Stop % Start Stop ID NO Site Site Sequence inhibition Site Site Linkage chemistry NO 433475 n/a n/a TCTTCAGCTTTCAGGCCAGC 17 443 462 sssssssssssssssssss 96 613027 5901 5920 GCCCTGCCGCTCGGCCGTCC 29 1 20 soooossssssssssooss 399 613028 5934 5953 TTCTCCTCCGGCCACTAGTG 22 34 53 soooossssssssssooss 400 613029 5987 6006 GAACGCGAGCCTCCCCAGGG 1 87 106 soooossssssssssooss 401 613030 6020 6039 GAGGCCGGCGGGCGGCGCAG 2 120 139 soooossssssssssooss 402 613031 6053 6072 GAGGGCGCGCGCCGGCGAAG 0 153 172 soooossssssssssooss 403 613032 6086 6105 TTGGTGCCGGAGCTGGTGGG 54 186 205 soooossssssssssooss 404 613033 6119 6138 GGCAGAAGGTGGGCGGTGGC 74 219 238 soooossssssssssooss 405 613034 6185 6204 TGATAGTCGACAGAGGCGAG 71 285 304 soooossssssssssooss 406 613035 73867 73886 CACTTCGAACTCCTGGCGGG 67 333 352 soooossssssssssooss 98 613036 73869 73888 ATCACTTCGAACTCCTGGCG 72 335 354 soooossssssssssooss 99 613037 73871 73890 CCATCACTTCGAACTCCTGG 69 337 356 soooossssssssssooss 100 613412 73879 73898 GTGATCTTCCATCACTTCGA 72 345 364 soooossssssssssooss 25 613038 73906 73925 CCTGTCCCCCAACCCGTACG 44 372 391 soooossssssssssooss 104 613039 73932 73951 TGCATGGTGTAGCCCCCCTG 80 398 417 soooossssssssssooss 117 613040 73934 73953 GGTGCATGGTGTAGCCCCCC 73 400 419 soooossssssssssooss 118 613041 73939 73958 GTCTTGGTGCATGGTGTAGC 77 405 424 soooossssssssssooss 407 613042 73958 73977 CGTCCGTGTCACCCTCTTGG 79 424 443 soooossssssssssooss 123 613044 83397 83416 CCTCAGATCCGTCCTCAGTG 67 n/a n/a soooossssssssssooss 408 613045 83430 83449 TGCTCTTAGCATCAGAGGTT 89 n/a n/a soooossssssssssooss 409 613046 85902 85921 CTAAGGGTGCTGTCACATCT 69 n/a n/a soooossssssssssooss 410 613047 85934 85953 GGCAGCCTGCTTGCCGGGAG 51 n/a n/a soooossssssssssooss 29 613048 85935 85954 CGGCAGCCTGCTTGCCGGGA 51 n/a n/a soooossssssssssooss 411 613049 85968 85987 TGGTTCCTTCTGGGATCTCC 76 n/a n/a soooossssssssssooss 412 613051 94719 94738 CTCGGAGGAAGCCTTCCTGG 53 n/a n/a soooossssssssssooss 413 613052 94752 94771 TGAGCTGGTGGCTCAGACCT 41 n/a n/a soooossssssssssooss 414 613053 94825 94844 CCTGTCCCCGAAGGTTGGCG 61 n/a n/a soooossssssssssooss 415 613054 94875 94894 GGTGCTTGAGCAGCTCAGGG 70 n/a n/a soooossssssssssooss 416 613055 94908 94927 GCGGCCCCTCCTGGTGCAGG 47 n/a n/a soooossssssssssooss 417 613056 94941 94960 CCGGCCTCTCTTTGCCCCCT 31 n/a n/a soooossssssssssooss 418 613057 94974 94993 CGTCGCGGTCTTCATCCACC 42 n/a n/a soooossssssssssooss 419 613058 95038 95057 GGCCGCCCATCTTGGGCTGG 0 n/a n/a soooossssssssssooss 420 613059 95071 95090 ATGCTGGTGGCTTCTCTGGC 53 n/a n/a soooossssssssssooss 421 613060 95098 95117 ATGGCACCCTCCGCTGGGAA 39 n/a n/a soooossssssssssooss 422 613061 95121 95140 AGAGGAAATCCACAGGGAGG 30 n/a n/a soooossssssssssooss 33 613062 95154 95173 GCTCTGAGGCTGGGATCTCT 70 n/a n/a soooossssssssssooss 423 613063 95187 95206 GCCCTTTGGCCCGCCCTACA 67 n/a n/a soooossssssssssooss 424 613064 95235 95254 CGTTGGGTGTGATTTCCACG 61 n/a n/a soooossssssssssooss 425 613065 95268 95287 AATGCTCCTCCGAGTGCGCC 85 n/a n/a soooossssssssssooss 426 613066 95313 95332 CCTCTGGCCCCTCTCCAGGG 43 n/a n/a soooossssssssssooss 427 613067 95346 95365 CTTTTGTGTCCTCTCCCAAA 42 n/a n/a soooossssssssssooss 428 613068 95379 95398 GCTGCTTTTCAGAGGGCTCT 51 n/a n/a soooossssssssssooss 429 613069 95412 95431 GGCTGACGGGCTTCCCCCGC 61 n/a n/a soooossssssssssooss 430 613070 95429 95448 TTTGAGTTGAGGGACCCGGC 33 n/a n/a soooossssssssssooss 431 613073 98559 98578 GCTTTTACTGACCATGCGAG 79 522 541 soooossssssssssooss 124 613074 98581 98600 CATCGCTTCCAGTCCCGTCT 78 544 563 soooossssssssssooss 134 613075 98587 98606 TTTTGTCATCGCTTCCAGTC 35 550 569 soooossssssssssooss 137 613076 101406 101425 AAGGTTTTAGCAGAGGAACG 63 n/a n/a soooossssssssssooss 37 613077 101457 101476 TCTGAGCTACCAGGAGTGGG 65 n/a n/a soooossssssssssooss 432 613078 101491 101510 GCACACAGCAGGGCTGGAGG 68 n/a n/a soooossssssssssooss 433 613079 101501 101520 GTGGCTCTGGGCACACAGCA 77 n/a n/a soooossssssssssooss 192 613080 101507 101526 AGGAAGGTGGCTCTGGGCAC 69 n/a n/a soooossssssssssooss 38 613081 101537 101556 GGGAAGTGACAGAAGAGACG 46 n/a n/a soooossssssssssooss 434 613082 101570 101589 GTTTCATCTCCTTTGCTCCA 57 n/a n/a soooossssssssssooss 435 613083 102995 103014 CCGCGGTGTGGCGATCTTCG 9 594 613 soooossssssssssooss 436 613084 103028 103047 GGCCTGGCCCTTCTGGCCTG 14 627 646 soooossssssssssooss 437 613085 103075 103094 GGTGTCTTTGGAGCGGGCGG 37 674 693 soooossssssssssooss 146 613086 105445 105464 CTTCTCTGGACTTGCTTAGT 45 n/a n/a soooossssssssssooss 438 613087 105475 105494 CTCTCAGATCTGGGCCCTGC 41 n/a n/a soooossssssssssooss 439 613088 105478 105497 CCTCTCTCAGATCTGGGCCC 49 n/a n/a soooossssssssssooss 440 613089 107932 107951 TAGCCGCTGCGATCCCCTGA 19 719 738 soooossssssssssooss 441 613090 107968 107987 CGGCTGCCGGGAGTGCCTGG 31 755 774 soooossssssssssooss 159 613091 108021 108040 CCACCTTCTTGGGCTCCCGG 17 808 827 soooossssssssssooss 442 613092 108040 108059 GGTGGAGTACGGACCACTGC 20 827 846 soooossssssssssooss 40 613093 108054 108073 AAGACGGCGACTTGGGTGGA 0 841 860 soooossssssssssooss 41 613094 108101 108120 CAGGTCTGGCATGGGCACGG 67 888 907 soooossssssssssooss 443 613095 108134 108153 CTCAGTGGAGCCGATCTTGG 31 921 940 soooossssssssssooss 199 613096 108167 108186 CACCTTCCCGCCTCCCGGCT 47 954 973 soooossssssssssooss 444 613097 121845 121864 GTTGCTAAGATCCAGCTTCT 49 n/a n/a soooossssssssssooss 445 613098 121865 121884 GAGCCACACTTGGACTGGAC 85 n/a n/a soooossssssssssooss 47 613099 121878 121897 GATATTATCCTTTGAGCCAC 88 n/a n/a soooossssssssssooss 446 613101 125784 125803 GGTCACCTTGCTCAGGTCAA 68 993 1012 soooossssssssssooss 447 613102 125817 125836 ATGATGGATGTTGCCTAATG 14 1026 1045 soooossssssssssooss 448 613103 130141 130160 ATTTTACTTCCACCTGGCCA 22 1057 1076 soooossssssssssooss 51

TABLE 12 Inhibition of Tau mRNA by 5-10-5 MOE gapmers with phosphorothioate and phosphodiester internucleoside linkages targeting SEQ ID NOs: 6 and 7 Target SEQ Target SEQ ISIS ID Start % ID NO NO Site Sequence inhibition Linkage chemistry NO 613043 6 442 GAGATTCTTTCAGGCCAGCG 17 soooossssssssssooss 449 613050 7 685 TTTCAGGCTCTTGGGTCACG 0 soooossssssssssooss 450 613071 7 1430 GCTTTGAGTTGAGGGACCCG 23 soooossssssssssooss 451 613072 7 1444 TACTGACCATGCGAGCTTTG 73 soooossssssssssooss 177 613100 7 2178 TATTTGCACACTGCCGCCTC 13 soooossssssssssooss 452

TABLE 13 Inhibition of Tau mRNA by 5-10-5 MOE gapmers with phosphorothioate and phosphodiester internucleoside linkages targeting SEQ ID NOs: 1 and 3 SEQ SEQ SEQ SEQ ID ID ID ID NO: 1 NO: 1 NO: 3 NO: 3 SEQ ISIS Start Stop % Start Stop ID NO Site Site Sequence inhibition Site Site Linkage chemistry NO 433475 n/a n/a TCTTCAGCTTTCAGGCCAGC 23 443 462 sssssssssssssssssss 96 613412 73879 73898 GTGATCTTCCATCACTTCGA 76 345 364 soooossssssssssooss 25 613104 130143 130162 AGATTTTACTTCCACCTGGC 59 1059 1078 soooossssssssssooss 453 613105 130176 130195 CGACTGGACTCTGTCCTTGA 47 1092 1111 soooossssssssssooss 454 613106 130225 130244 TATTTCCTCCGCCAGGGACG 90 1141 1160 soooossssssssssooss 455 613107 135483 135502 CGCGGAAGGTCAGCTTGTGG 43 1174 1193 soooossssssssssooss 456 613108 135516 135535 CCGCCCCGTGGTCTGTCTTG 44 1207 1226 soooossssssssssooss 457 613109 135549 135568 CCCCAGACACCACTGGCGAC 36 1240 1259 soooossssssssssooss 458 613110 135590 135609 GCCGGTGGAGGAGACATTGC 51 1281 1300 soooossssssssssooss 459 613111 135654 135673 CCAGGGAGGCAGACACCTCG 61 1345 1364 soooossssssssssooss 245 613112 135697 135716 AATTATTGACCGCCCCAGGG 33 1388 1407 soooossssssssssooss 460 613113 135730 135749 TTTTTCCACACTCTCTCATT 41 1421 1440 soooossssssssssooss 461 613114 135783 135802 AACTGCGAGGAGCAGCTGGG 51 1474 1493 soooossssssssssooss 462 613115 135786 135805 CCGAACTGCGAGGAGCAGCT 78 1477 1496 soooossssssssssooss 248 613116 135816 135835 CAAAAGCAGGTTAAGTGATT 28 1507 1526 soooossssssssssooss 463 613117 135849 135868 CTGATTTTGAAGTCCCGAGC 69 1540 1559 soooossssssssssooss 464 613118 135869 135888 ATTTGCTCTTACTCCCATCA 80 1560 1579 soooossssssssssooss 465 613119 135882 135901 TTGGAAAGATGAAATTTGCT 52 1573 1592 soooossssssssssooss 466 613120 135948 135967 TGTTGGATGTGGCCATGTTT 63 1639 1658 soooossssssssssooss 467 613121 136049 136068 GTCCCTTGAAATCCCCCAGA 45 1740 1759 soooossssssssssooss 468 613122 136082 136101 CCACAACAGGGCCAGAGGTG 55 1773 1792 soooossssssssssooss 469 613123 136115 136134 TCCTTTGTTGCTGCCACTGC 59 1806 1825 soooossssssssssooss 470 613124 136148 136167 GCCTGTGGCTCCACGAACAC 72 1839 1858 soooossssssssssooss 471 613125 136181 136200 CCCCGTCACACTCACACAAG 43 1872 1891 soooossssssssssooss 472 613126 136214 136233 GGCCTCCCCCGTGGCCTCCC 37 1905 1924 soooossssssssssooss 473 613127 136247 136266 TGCTTCCTCTCCCCTCTGCC 41 1938 1957 soooossssssssssooss 474 613128 136295 136314 GATGTCTACTCTCCAGCACG 32 1986 2005 soooossssssssssooss 475 613129 136328 136347 ATAGGCCTTGGCTCTCCCAG 73 2019 2038 soooossssssssssooss 476 613130 136361 136380 AGGACAGGCGGCCGCTCAGA 46 2052 2071 soooossssssssssooss 477 613131 136394 136413 TGACCCACAGCAGGCCCCCA 14 2085 2104 soooossssssssssooss 478 613132 136427 136446 CTCCCACAGGCTGCCCTGCA 46 2118 2137 soooossssssssssooss 479 613133 136460 136479 CAGCTTGCCTTCTCTTTTTA 58 2151 2170 soooossssssssssooss 480 613134 136491 136510 GGAGGTCATCCACGAAGTGC 45 2182 2201 soooossssssssssooss 61 613135 136493 136512 AAGGAGGTCATCCACGAAGT 41 2184 2203 soooossssssssssooss 481 613136 136515 136534 GACATCAAGGTCAGTCTTTT 80 2206 2225 soooossssssssssooss 482 613137 136541 136560 AGGGAGGAAGAGGCCAGCGC 60 2232 2251 soooossssssssssooss 483 613138 136574 136593 AGCCCCTCAACTCAGGCCCC 53 2265 2284 soooossssssssssooss 484 613139 136607 136626 TCAATAAAACAGGGTTTCTG 38 2298 2317 soooossssssssssooss 485 613140 136640 136659 CAAAATCATGGCAGCAGTTC 26 2331 2350 soooossssssssssooss 486 613141 136673 136692 GTTAGCCCTAAAGTCCCAGG 76 2364 2383 soooossssssssssooss 487 613142 136706 136725 CCCAAGAGGCACAAGTCCTT 57 2397 2416 soooossssssssssooss 488 613143 136739 136758 CAGTGGCCCAGGCTTGGAAA 63 2430 2449 soooossssssssssooss 489 613144 136748 136767 CAGAGATGCCAGTGGCCCAG 43 2439 2458 soooossssssssssooss 67 613145 136772 136791 TGCCTCCCAGACCCCCACAC 23 2463 2482 soooossssssssssooss 490 613146 136805 136824 AGTGGCCGTGGGAAGGACAG 17 2496 2515 soooossssssssssooss 491 613147 136838 136857 ACAACAGCACAGCGGCGCAG 61 2529 2548 soooossssssssssooss 492 613148 136845 136864 ACGGCAGACAACAGCACAGC 59 2536 2555 soooossssssssssooss 493 613149 136899 136918 GAATTCGGGACATTGTGACG 42 2590 2609 soooossssssssssooss 494 613150 136933 136952 CAGGGTCATTACTGAGAAGG 39 2624 2643 soooossssssssssooss 495 613151 136966 136985 CTCAGTATGGAGTAGGTACC 52 2657 2676 soooossssssssssooss 496 613152 136999 137018 TGTGCCTGGACTTTGCCTTC 61 2690 2709 soooossssssssssooss 497 613153 137032 137051 GGAACTGAGAGTGAGAGGCT 62 2723 2742 soooossssssssssooss 498 613154 137077 137096 GAATCAGATCATGAGATTCG 30 2768 2787 soooossssssssssooss 499 613155 137110 137129 CACATCTGTGACGGGAGGAG 26 2801 2820 soooossssssssssooss 500 613156 137143 137162 CACCTAGGGTCACAGCTGAG 33 2834 2853 soooossssssssssooss 501 613157 137176 137195 GAAAGGGCTCTCTCCATGTC 54 2867 2886 soooossssssssssooss 502 613158 137214 137233 GTGGGCTCAGCACAGGAAGG 23 2905 2924 soooossssssssssooss 503 613159 137229 137248 ACCCAGCCTGCTGCTGTGGG 36 2920 2939 soooossssssssssooss 504 613160 137247 137266 ACCACTGACAACCAAGACAC 22 2938 2957 soooossssssssssooss 505 613161 137280 137299 GCCCTGGGTGCCTTGCCCTT 53 2971 2990 soooossssssssssooss 506 613162 137322 137341 ACAAGCTAGGGTGCAAGTGG 47 3013 3032 soooossssssssssooss 507 613163 137364 137383 GGAGCTGAGCAGCGGGCTGG 45 3055 3074 soooossssssssssooss 508 613164 137401 137420 CCCTTTGTCGGGTGTGGAGG 8 3092 3111 soooossssssssssooss 509 613165 137453 137472 TGGCTTCCAGCTGGGACTGG 65 3144 3163 soooossssssssssooss 510 613166 137486 137505 ATATGTTCAGCTGCTCCAGC 58 3177 3196 soooossssssssssooss 511 613167 137529 137548 ACTCAACAGGGTGCAGATGG 54 3220 3239 soooossssssssssooss 512 613168 137562 137581 GAATCCAAGCATAAACAGAC 55 3253 3272 soooossssssssssooss 513 613169 137595 137614 TTTTTTTCTTTTCACTATCA 34 3286 3305 soooossssssssssooss 514 613170 137629 137648 AGCATTTCAAGATACATGCG 59 3320 3339 soooossssssssssooss 515 613171 137692 137711 CACACGAGTCCCAGTGTGGG 11 3383 3402 soooossssssssssooss 516 613172 137761 137780 TTGGGTCCCAGGTGCTGAGG 40 3452 3471 soooossssssssssooss 517 613173 137794 137813 ACGGCCTCCTTAGCTGCTAG 73 3485 3504 soooossssssssssooss 518 613174 137827 137846 TAATCCTGTGCTTCAGGCCT 32 3518 3537 soooossssssssssooss 519 613175 137881 137900 TGACACAGGGAGCCCCAAGG 40 3572 3591 soooossssssssssooss 520 613176 137914 137933 AGCCAGACCAGCCACAAGAC 16 3605 3624 soooossssssssssooss 521 613177 137947 137966 GCTATGACCAGAGAGAACCA 29 3638 3657 soooossssssssssooss 522 613178 137967 137986 GGACTGCCATGAGACTTCGG 26 3658 3677 soooossssssssssooss 523 613179 137980 137999 TAAGCCTCCTTTGGGACTGC 64 3671 3690 soooossssssssssooss 524 613180 138013 138032 AGTGGCTTCCTTTTTCTTGT 69 3704 3723 soooossssssssssooss 525

TABLE 14 Inhibition of Tau mRNA by 5-10-5 MOE gapmers with phosphorothioate and phosphodiester internucleoside linkages targeting SEQ ID NOs: 1 and 3 SEQ SEQ SEQ SEQ ID ID ID ID NO: 1 NO: 1 NO: 3 NO: 3 SEQ ISIS Start Stop % Start Stop ID NO Site Site Sequence inhibition Site Site Linkage chemistry NO 433475 n/a n/a TCTTCAGCTTTCAGGCCAGC 23 433 452 sssssssssssssssssss 96 613258 18371 18390 GGCAATGCCTGGAAGAAACT 75 n/a n/a soooossssssssssooss 526 613259 18411 18430 CGCTGGTGAGAGTGTGACTG 69 n/a n/a soooossssssssssooss 527 613260 18449 18468 ATCCGAGGGTGAGATGATAA 75 n/a n/a soooossssssssssooss 528 613261 19611 19630 AATAATTTCTCCAGGCTCAT 62 n/a n/a soooossssssssssooss 529 613262 20955 20974 GCTGACTTCGATTTGTTATT 84 n/a n/a soooossssssssssooss 530 613263 22139 22158 TATCATATGGCACAAATTCT 32 n/a n/a soooossssssssssooss 531 613264 23662 23681 CTACCTGGGACCACACCTTA 61 n/a n/a soooossssssssssooss 532 613265 25046 25065 GGGTGGAGAGCCTGATGGTC 47 n/a n/a soooossssssssssooss 533 613266 25418 25437 CTGTGAGCATCATGAAAGCC 56 n/a n/a soooossssssssssooss 534 613267 25606 25625 TTTTATTATGAGGGAGATCA 48 n/a n/a soooossssssssssooss 535 613268 26186 26205 AATAGTAGAGAAATGGAAAT 39 n/a n/a soooossssssssssooss 536 613269 26690 26709 TAACATGACAGGGTTAGGCC 67 n/a n/a soooossssssssssooss 537 613270 26917 26936 GCCCACACACATGCTCATGC 57 n/a n/a soooossssssssssooss 538 613271 27160 27179 TGTGTTCAGTTACATGCCTT 82 n/a n/a soooossssssssssooss 539 613272 27569 27588 GACCACCATGGTGGGCTGAC 39 n/a n/a soooossssssssssooss 540 613273 28227 28246 GGCACCTCTCTGCAGACTTT 85 n/a n/a soooossssssssssooss 541 613274 30102 30121 TGCTTCACACACCACAATGG 60 n/a n/a soooossssssssssooss 542 613275 31769 31788 AACATTGCCGAGCACCAGCC 63 n/a n/a soooossssssssssooss 543 613276 31832 31851 GACTCAGGAACATGTTAGCT 70 n/a n/a soooossssssssssooss 544 613277 32045 32064 CACGGCTCAGCAGAGGAGAC 23 n/a n/a soooossssssssssooss 545 613278 32324 32343 GCTGCTATCCTTCCTGGGCC 57 n/a n/a soooossssssssssooss 546 613279 32760 32779 GGAAATGAAAGCTTCCCACG 61 n/a n/a soooossssssssssooss 547 613280 33359 33378 AACAGTCAGAATACCCCCAA 72 n/a n/a soooossssssssssooss 548 613281 34056 34075 TATAGGTAGAATATAAAACT 17 n/a n/a soooossssssssssooss 549 613282 35266 35285 TATGAACAAAACTGCAGGAC 45 n/a n/a soooossssssssssooss 550 613283 35839 35858 AGCCTCCAGAGTGACAGGTG 63 n/a n/a soooossssssssssooss 551 613284 36223 36242 CCATAAGCCACTCCTGGTTG 0 n/a n/a soooossssssssssooss 552 613285 36410 36429 CAGCTTGGAAGATACAGGAG 63 n/a n/a soooossssssssssooss 553 613286 38647 38666 CAAACTTAAGAAAAGTTGCA 67 n/a n/a soooossssssssssooss 554 613287 39104 39123 TGCTGGCAAAGAAGACAGGA 69 n/a n/a soooossssssssssooss 555 613288 40506 40525 CCACTGATGGCCGGGTGCAG 27 n/a n/a soooossssssssssooss 556 613289 42727 42746 TGGGTGTCAAATGGCTGGCT 55 n/a n/a soooossssssssssooss 557 613290 42955 42974 TAGAAAGCTATTTAATAAAT 0 n/a n/a soooossssssssssooss 558 613291 43109 43128 AGCCGTACATCAATGGTACA 71 n/a n/a soooossssssssssooss 559 613292 46627 46646 GGCCCTACCATGGTCGATTT 57 n/a n/a soooossssssssssooss 560 613293 47399 47418 ATCAGTGCCAGAGACAGAGG 5 n/a n/a soooossssssssssooss 561 613294 48115 48134 TATTATTAACGAACAAAAAA 0 n/a n/a soooossssssssssooss 562 613295 48223 48242 AGAGCAATACTTTGTGCTTA 51 n/a n/a soooossssssssssooss 563 613296 48608 48627 AGACTCCTCAAATGTGCGCC 53 n/a n/a soooossssssssssooss 564 613297 49204 49223 GAATGACGGCCGCCTCCTCT 39 n/a n/a soooossssssssssooss 565 613298 49696 49715 TATGGAGGGCTTCTGGCCTC 18 n/a n/a soooossssssssssooss 566 613299 51161 51180 GAGAAAGAGCGAGTAATTAG 44 n/a n/a soooossssssssssooss 567 613300 52250 52269 TAGCTAGAAACTGTGTCCCT 41 n/a n/a soooossssssssssooss 568 613301 53908 53927 CTGTGAGATCATCCCCTGGT 51 n/a n/a soooossssssssssooss 569 613302 54457 54476 CAGCGCAGTGCAGGGAGTCA 41 n/a n/a soooossssssssssooss 570 613303 55975 55994 GGGTACACAGAGAGGCCACT 53 n/a n/a soooossssssssssooss 571 613304 56155 56174 GAACGATGCACTGCCTTGGC 54 n/a n/a soooossssssssssooss 572 613305 57402 57421 CACCATATTACACCAGTTGT 60 n/a n/a soooossssssssssooss 573 613306 57696 57715 CTGAAATGAATCCCCTGAAC 0 n/a n/a soooossssssssssooss 574 613307 57733 57752 GTGTCCACGAATGCCCCTGG 40 n/a n/a soooossssssssssooss 575 613308 58392 58411 ACGGATGGTTGCTGCTGATG 47 n/a n/a soooossssssssssooss 576 613309 61917 61936 GTCCCTTTTGCTCCCAAAGA 18 n/a n/a soooossssssssssooss 577 613310 62489 62508 CAGTCTTTTAATAAGTGAAA 50 n/a n/a soooossssssssssooss 578 613311 63219 63238 TTAGCTAAATTGAGGCCCGG 31 n/a n/a soooossssssssssooss 579 613312 63257 63276 GGAAAGGTATTGCCTCCTGA 42 n/a n/a soooossssssssssooss 580 613313 63627 63646 GGTGACTACGCAGCCTGGAC 57 n/a n/a soooossssssssssooss 581 613314 64559 64578 TTCAGGGCAATCCTTGAGGT 41 n/a n/a soooossssssssssooss 582 613315 64872 64891 TGCTAATGCTTTGGGACCTA 44 n/a n/a soooossssssssssooss 583 613316 66922 66941 TGGGTAGAGAGCAACTGTTC 32 n/a n/a soooossssssssssooss 584 613317 67104 67123 ACACCCCTTCTCTGCACATC 25 n/a n/a soooossssssssssooss 585 613318 68893 68912 CTGCTAGAGTTCGCCAACAG 59 n/a n/a soooossssssssssooss 586 613319 69457 69476 TCTTATTCTCTAGGTGGGCT 70 n/a n/a soooossssssssssooss 587 613320 70922 70941 CAGTGCAAGTGGGTATGAAG 30 n/a n/a soooossssssssssooss 588 613321 70928 70947 CAGATGCAGTGCAAGTGGGT 44 n/a n/a soooossssssssssooss 589 613322 71310 71329 GAAAAACTTTGTCCATAATT 41 n/a n/a soooossssssssssooss 590 613323 71665 71684 TAATGAATGTATAACAGAAA 0 n/a n/a soooossssssssssooss 591 613324 71702 71721 AAAAGGCAGATACTTTGTGG 56 n/a n/a soooossssssssssooss 592 613325 72456 72475 CAGAGGTGTGAAAGTAAAAG 0 n/a n/a soooossssssssssooss 593 613326 72566 72585 CAGCTAAAGCTTTGTGAACA 31 n/a n/a soooossssssssssooss 594 613327 72612 72631 TGCAGCCCAGGTGAGCCAGC 49 n/a n/a soooossssssssssooss 595 613328 73748 73767 GCCTCTCCTCCCAGTTCATG 12 n/a n/a soooossssssssssooss 596 613329 75140 75159 GTTTTCTTATTTTTAGCATT 87 n/a n/a soooossssssssssooss 597 613330 76340 76359 GGAGGGACGAGAAACAGCAG 0 n/a n/a soooossssssssssooss 598 613331 79041 79060 AGCTTCCAGCATGAACATAG 77 n/a n/a soooossssssssssooss 599 613332 79971 79990 GAGCCCTGATATGATAGAGG 57 n/a n/a soooossssssssssooss 600 613333 80360 80379 TGTACCATGCCTGGCAGATG 78 n/a n/a soooossssssssssooss 601 613334 81325 81344 GTTAGATGTATGTATGATGG 0 n/a n/a soooossssssssssooss 602 613412 73879 73898 GTGATCTTCCATCACTTCGA 80 345 364 soooossssssssssooss 25

TABLE 15 Inhibition of Tau mRNA by 5-10-5 MOE gapmers with phosphorothioate and phosphodiester internucleoside linkages targeting SEQ ID NOs: 1 and 3 SEQ SEQ SEQ SEQ ID ID ID ID NO: 1 NO: 1 NO: 3 NO: 3 SEQ ISIS Start Stop % Start Stop ID NO Site Site Sequence inhibition Site Site Linkage chemistry NO 433475 n/a n/a TCTTCAGCTTTCAGGCCAGC 31 443 462 sssssssssssssssssss 96 613335 82136 82155 GCCTCCTCCTAGAGCTCCTG 81 n/a n/a soooossssssssssooss 603 613336 83231 83250 CGGCCTCGCAACAAACCCAC 55 n/a n/a soooossssssssssooss 604 613337 83820 83839 TCCACTGACCTGCCCCTTCC 66 n/a n/a soooossssssssssooss 605 613338 84269 84288 TTTGCAACTTGTTCCTGCTT 84 n/a n/a soooossssssssssooss 606 613339 85073 85092 TCTTGGGTGCAGGCGGAGCA 53 n/a n/a soooossssssssssooss 607 613340 86117 86136 ACCCAAGATTCCCAGGAGCC 77 n/a n/a soooossssssssssooss 608 613341 86191 86210 AGGCCCTAAATGCTCTGAGA 75 n/a n/a soooossssssssssooss 609 613342 86251 86270 AGAAGTCTCTGGAACTGACG 70 n/a n/a soooossssssssssooss 610 613343 86714 86733 TCACCAAACCAATGGCTGGC 67 n/a n/a soooossssssssssooss 611 613344 86774 86793 GGTGAAATCCTGGCTAGGCC 89 n/a n/a soooossssssssssooss 612 613345 88487 88506 TGCAGATTCGGCCTGAGTTT 72 n/a n/a soooossssssssssooss 613 613346 89131 89150 CCAAACAAGAAAGACTGAGA 55 n/a n/a soooossssssssssooss 614 613347 89994 90013 CCATGTCCCTCCCCCCAACC 12 n/a n/a soooossssssssssooss 615 613348 90126 90145 ATTAGAAAAAAACCACTAGG 37 n/a n/a soooossssssssssooss 616 613349 92246 92265 AGCTTACTAGAGTGCTCTTG 81 n/a n/a soooossssssssssooss 617 613350 92915 92934 ATTTTTTTCTGGTTCATTCA 71 n/a n/a soooossssssssssooss 618 613351 94316 94335 CGCTTAAGAGAGGGAGCATC 72 n/a n/a soooossssssssssooss 619 613352 94673 94692 GTGGAGATACGCAGTGGTGG 68 n/a n/a soooossssssssssooss 620 613353 95790 95809 ACACCATGAGGGCACCCGTC 82 n/a n/a soooossssssssssooss 35 613354 95918 95937 CCCAAACCTGGCTCTGTCAC 65 n/a n/a soooossssssssssooss 621 613355 97224 97243 ATCAACCAGCACCATCCACG 65 n/a n/a soooossssssssssooss 622 613356 97314 97333 GGGTCTGTAAGCCTAAAGTG 53 n/a n/a soooossssssssssooss 623 613357 97315 97334 TGGGTCTGTAAGCCTAAAGT 56 n/a n/a soooossssssssssooss 624 613358 97368 97387 TATACTATGTGGCAGAATCA 48 n/a n/a soooossssssssssooss 625 613359 97478 97497 CACATTCCCTCTACCCCATG 57 n/a n/a soooossssssssssooss 626 613360 97507 97526 TCATAGATTTTCTTCTTTGG 75 n/a n/a soooossssssssssooss 627 613361 98881 98900 ACACACCTTCATTTACTGTC 95 n/a n/a soooossssssssssooss 313 98918 98937 n/a n/a soooossssssssssooss 613362 98882 98901 AACACACCTTCATTTACTGT 83 n/a n/a soooossssssssssooss 314 98919 98938 n/a n/a soooossssssssssooss 613363 98883 98902 AAACACACCTTCATTTACTG 79 n/a n/a soooossssssssssooss 315 98920 98939 n/a n/a soooossssssssssooss 613364 98884 98903 CAAACACACCTTCATTTACT 63 n/a n/a soooossssssssssooss 316 98921 98940 n/a n/a soooossssssssssooss 613365 98885 98904 TCAAACACACCTTCATTTAC 59 n/a n/a soooossssssssssooss 317 98922 98941 n/a n/a soooossssssssssooss 613366 98886 98905 TTCAAACACACCTTCATTTA 67 n/a n/a soooossssssssssooss 628 98923 98942 n/a n/a soooossssssssssooss 613367 98887 98906 TTTCAAACACACCTTCATTT 72 n/a n/a soooossssssssssooss 318 98924 98943 n/a n/a soooossssssssssooss 613368 98888 98907 TTTTCAAACACACCTTCATT 59 n/a n/a soooossssssssssooss 319 98925 98944 n/a n/a soooossssssssssooss 613369 98889 98908 GTTTTCAAACACACCTTCAT 90 n/a n/a soooossssssssssooss 320 98926 98945 n/a n/a soooossssssssssooss 613370 98890 98909 GGTTTTCAAACACACCTTCA 96 n/a n/a soooossssssssssooss 321 98927 98946 n/a n/a soooossssssssssooss 613371 98891 98910 TGGTTTTCAAACACACCTTC 95 n/a n/a soooossssssssssooss 322 98928 98947 n/a n/a soooossssssssssooss 613372 99560 99579 CCCCAGCTGGCCGAGGCCCA 81 n/a n/a soooossssssssssooss 629 613373 99816 99835 GAGAAGGGTCCTGTCCCAGA 76 n/a n/a soooossssssssssooss 630 613374 100743 100762 AGTAAGTTGATGGAGATCAT 83 n/a n/a soooossssssssssooss 631 613375 102894 102913 CACCAGCCCCACTCCGCCAC 61 n/a n/a soooossssssssssooss 632 613376 103875 103894 TGCTCTGTCTCCAGAGACAT 70 n/a n/a soooossssssssssooss 633 613377 104615 104634 ATCCAAGATCCAGGCCAGGC 63 n/a n/a soooossssssssssooss 634 613378 105870 105889 AGAGACCCAGGGTGACCAGT 76 n/a n/a soooossssssssssooss 635 613379 106909 106928 CAAATTTCTCGATCCCCTTC 18 n/a n/a soooossssssssssooss 636 613380 108411 108430 CCTGAGGAGGGCACTCACAG 61 n/a n/a soooossssssssssooss 637 613381 108891 108910 GTGGCTACCCACGGCCACAG 1 n/a n/a soooossssssssssooss 638 613382 108939 108958 CTACCAGGAGCACACACAGA 65 n/a n/a soooossssssssssooss 639 613383 109303 109322 CCCTAAGCCATGGTGGGTTT 72 n/a n/a soooossssssssssooss 640 613384 109995 110014 CCCTTGTCCCCACAGCCACA 72 n/a n/a soooossssssssssooss 641 613385 110198 110217 GGGTCACCTGTGAGTTATTT 62 n/a n/a soooossssssssssooss 642 613386 113567 113586 AGCTGCCTCCAGTTGTTTAT 85 n/a n/a soooossssssssssooss 643 613387 114056 114075 ACTTTAGCTCCATCTTCTCA 49 n/a n/a soooossssssssssooss 644 613388 114755 114774 CTATAATGGTCAGTGGTGTA 67 n/a n/a soooossssssssssooss 645 613389 115899 115918 TGAAGGAGAGGCTGTGGTGA 41 n/a n/a soooossssssssssooss 646 613390 117387 117406 TGTGACAACAAAGTTGTCCT 46 n/a n/a soooossssssssssooss 647 613391 117654 117673 CCCTGCAATCAACACAGGAG 65 n/a n/a soooossssssssssooss 648 613392 119050 119069 TCTAACATGCACATATTTAC 60 n/a n/a soooossssssssssooss 649 613393 119190 119209 AGCACTGGGTGTTTACAACA 84 n/a n/a soooossssssssssooss 650 613394 119594 119613 AGAAGTGATATCATATCCTA 85 n/a n/a soooossssssssssooss 324 119905 119924 n/a n/a soooossssssssssooss 613395 119595 119614 AAGAAGTGATATCATATCCT 68 n/a n/a soooossssssssssooss 325 119906 119925 n/a n/a soooossssssssssooss 613396 120782 120801 ACCCGGTGACCTTTCCTCTC 67 n/a n/a soooossssssssssooss 651 613397 121728 121747 GAGTGACATGCGCCACCCTG 87 n/a n/a soooossssssssssooss 203 613398 121794 121813 AAAAGGATGAGTGACACGCC 66 n/a n/a soooossssssssssooss 205 613399 121940 121959 GCCACAGCACGGCGCATGGG 89 n/a n/a soooossssssssssooss 652 613400 124954 124973 GCACCGCTCCCTCTGGGAAT 49 n/a n/a soooossssssssssooss 653 613401 125298 125317 AAATCTTGCTGGAAAGGCAG 64 n/a n/a soooossssssssssooss 654 613402 125428 125447 GACCCAGACATTTGCTCAGC 86 n/a n/a soooossssssssssooss 655 613403 125982 126001 GGGCTCCCGCAAGTTTCACA 71 n/a n/a soooossssssssssooss 656 613404 127460 127479 GAGAGGCCCTGTGGACAGCT 75 n/a n/a soooossssssssssooss 657 613405 130381 130400 CCCACTGGATGCTGCTGAGG 34 n/a n/a soooossssssssssooss 658 613406 131662 131681 GTGCCTAGTAACCCATTTAG 54 n/a n/a soooossssssssssooss 659 613407 131796 131815 CACCCACAGCAAGACCTAGA 49 n/a n/a soooossssssssssooss 660 613408 133056 133075 ACACAGAGAAGCTTCCAGAG 70 n/a n/a soooossssssssssooss 661 613409 133304 133323 AGCTATCACATGGACGAGTT 67 n/a n/a soooossssssssssooss 662 613410 133548 133567 CATTTAATACAGGCTTTGAT 38 n/a n/a soooossssssssssooss 663 613411 134599 134618 AGCCACTGATGCCCAGACAT 69 n/a n/a soooossssssssssooss 664 613412 73879 73898 GTGATCTTCCATCACTTCGA 82 345 364 soooossssssssssooss 25

TABLE 16 Inhibition of Tau mRNA by 5-10-5 MOE gapmers with phosphorothioate and phosphodiester internucleoside linkages targeting SEQ ID NOs: 5 and 6 Target SEQ Target SEQ ISIS ID Start % ID NO NO Site Sequence inhibition Linkage chemistry NO 613239 5 3 TGCCCTTCGCGGTCCCTTCG 11 soooossssssssssooss 396 613241 5 424 CTGTCCCCCAAACCCGTACG 48 soooossssssssssooss 397 613248 6 524 GCTTCCGCTGTTGGAGTGCT 59 soooossssssssssooss 398

Example 8 Dose-Dependent Antisense Inhibition of Human Tau in SH-SY5Y Cells by 5-10-5 MOE Gapmers with Phosphorothioate and Phosphodiester Internucleoside Linkages

Gapmers from studies described above exhibiting significant in vitro inhibition of tau mRNA were selected and tested at various doses in SH-SY-5Y cells. Cells were plated at a density of 20,000 cells per well and transfected using electroporation with 1.25 μM, 2.500 μM, 5.00 μM, 10.00 μM, and 20.00 μM concentrations of antisense oligonucleotide, as specified in the Table below. After a treatment period of approximately 16 hours, RNA was isolated from the cells and Tau mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3104 was used to measure mRNA levels. Tau mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of Tau, relative to untreated control cells. Tau mRNA levels were significantly reduced in a dose-dependent manner in antisense oligonucleotide treated cells.

TABLE 17 ISIS No 1.25 μM 2.50 μM 5.00 μM 10.00 μM 20.00 μM 613118 46 47 73 92 93 613136 36 70 83 88 92 613115 32 49 82 86 94 613329 73 80 92 94 92 613273 46 24 48 60 85 613262 31 72 76 87 94 613271 44 66 84 89 94 613370 92 92 95 98 98 613361 88 86 89 94 96 613371 80 88 91 95 97 613369 58 85 89 95 96 613399 61 77 90 94 96 613344 80 79 93 95 95 613397 64 74 79 93 95 613412 45 66 70 81 96

TABLE 18 ISIS No 1.25 μM 2.50 μM 5.00 μM 10.00 μM 20.00 μM 613255 54 61 80 91 90 613065 22 61 62 83 89 613257 45 69 82 86 91 613098 70 57 83 92 98 613253 44 67 74 86 91 613039 21 77 85 90 95 613188 0 35 72 86 87 613042 33 45 73 90 94 613233 21 54 69 75 89 613073 59 67 83 92 97 613187 8 54 74 61 84 613402 10 58 62 81 87 613045 57 72 92 96 95 613412 20 52 67 84 90 613099 71 66 89 91 95

Example 9 Antisense Inhibition of Human Tau in SH-SY5Y Cells by 5-10-5 MOE, 5-8-5 MOE, 4-8-6 MOE, or 6-8-4 MOE Gapmers

Antisense oligonucleotides were designed targeting a tau nucleic acid and were tested for their effects on Tau mRNA in vitro. The antisense oligonucleotides were tested in a series of experiments that had similar culture conditions. ISIS 613412 was also included in the assays. The results for each experiment are presented in separate tables shown below. Cultured SH-SY5Y cells at a density of 20,000 cells per well were transfected using electroporation with 8,000 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and Tau mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3104 was used to measure mRNA levels. Tau mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®). Results are presented as percent inhibition of tau, relative to untreated control cells.

The newly designed chimeric antisense oligonucleotides in the Tables below were designed as 5-8-5 MOE, 4-8-6 MOE, or 6-8-4 MOE gapmers. The 5-8-5 MOE gapmers are 18 nucleosides in length, wherein the central gap segment comprises of eight 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five nucleosides each. The 4-8-6 MOE gapmers are 18 nucleosides in length, wherein the central gap segment comprises of eight 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising four and six nucleosides respectively. The 6-8-4 MOE gapmers are 18 nucleosides in length, wherein the central gap segment comprises of eight 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising six and four nucleosides respectively. Each nucleoside in the 5′ wing segment and each nucleoside in the 3′ wing segment has a 2′-MOE modification. The internucleoside linkage motif throughout for each gapmer in the tables below, except for ISIS 613412, is 5′-sooosssssssssooss-3′, wherein each “s” represents a phosphorothioate internucleoside linkage and wherein each “o” represents a phosphodiester internucleoside linkage. The internucleoside linkage motif for ISIS 613412 is 5′-soooossssssssssooss-3′, wherein each “s” represents a phosphorothioate internucleoside linkage and wherein each “o” represents a phosphodiester internucleoside linkage. All cytosine residues throughout each gapmer are 5-methylcytosines. “Start site” indicates the 5′-most nucleoside to which the gapmer is targeted in the human gene sequence. “Stop site” indicates the 3′-most nucleoside to which the gapmer is targeted human gene sequence. Each gapmer listed in the Tables below is targeted to either SEQ ID NO: 1 (GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to 9381000), SEQ ID NO: 4 (GENBANK Accession No. NT_010783.14 truncated from nucleotides 2624000 to 2761000), SEQ ID NO: 5 (GENBANK Accession No. DR002467.1), or SEQ ID NO: 6 (GENBANK Accession No. NM_001203251.1). ‘n/a.’ indicates that the antisense oligonucleotide does not target that particular gene sequence with 100% complementarity.

TABLE 19 Inhibition of tau mRNA by 5-10-5 MOE and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 and SEQ ID NO: 4 SEQ SEQ SEQ SEQ ID ID ID ID NO: 1 NO: 1 NO: 4 NO: 4 SEQ ISIS Start Stop % Start Stop ID NO Site Site Sequence inhibition Site Site Motif NO 613412 73879 73898 GTGATCTTCCATCACTTCGA 83 69842 69861 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 89 94855 94872 5-8-5 665 98928 98945 94892 94909 621197 n/a n/a TCGCCAACAGACATGTGA 39 64850 64867 5-8-5 666 621196 67898 67915 TGAAGGAGCCTGGGAAGT 15 63891 63908 5-8-5 667 621198 69079 69096 AGCAGAGATGTGGCTGGG 50 65044 65061 5-8-5 668 621199 69622 69639 GTGAGGTTGAAAAGTTTC 34 65587 65604 5-8-5 669 621200 69802 69819 TGGTTTCTGATGGTTTTT 43 65767 65784 5-8-5 670 621201 70023 70040 GTTGGAACTCCTGGGATC 27 65988 66005 5-8-5 671 621202 70259 70276 TCAGCATTTTTCTCCTCT 47 66224 66241 5-8-5 672 621203 70531 70548 CTGGAATGGTACGATCTC 52 66496 66513 5-8-5 673 621204 70750 70767 CCACTCTGGTGCAGGACG 29 66715 66732 5-8-5 674 621205 70930 70947 CAGATGCAGTGCAAGTGG 43 66895 66912 5-8-5 675 621206 71110 71127 GGGTTTCACCAGACCTCA 29 67075 67092 5-8-5 676 621207 71306 71323 CTTTGTCCATAATTTTTT 34 67271 67288 5-8-5 677 621208 71487 71504 TATAGCATGAGCATTTGT 42 67452 67469 5-8-5 678 621209 71667 71684 TAATGAATGTATAACAGA 2 67632 67649 5-8-5 679 621210 71850 71867 TCCTCCTGGCTGCCCTGT 33 67815 67832 5-8-5 680 621211 72030 72047 CATCCTGCCATACCAGAC 32 67995 68012 5-8-5 681 621212 72224 72241 TATCCGCCTCTCAAAGTG 0 68189 68206 5-8-5 682 621213 72410 72427 CACATTTGCATATTTTCA 65 68373 68390 5-8-5 683 621214 72590 72607 TCGGGTTGCCCCATTCAA 27 68553 68570 5-8-5 684 621215 72770 72787 AACCACCATTTACTCACA 35 68733 68750 5-8-5 685 621216 72950 72967 ACCGGCACCCACCAGGTC 22 68913 68930 5-8-5 686 621217 73130 73147 AGGGACAGCATCAGCAGA 22 69093 69110 5-8-5 687 621218 73437 73454 AGGCATGTGGCAGGTGCC 12 69400 69417 5-8-5 688 621219 73617 73634 ACCAACACCGCAGCAGTT 35 69580 69597 5-8-5 689 621220 73797 73814 GTTCTGAGGAGTGTTGGG 0 69760 69777 5-8-5 690 621221 73978 73995 ACTAACCTTTCAGGCCAG 33 69941 69958 5-8-5 691 621222 74273 74290 TAAATTATTTTAGAGACG 0 70236 70253 5-8-5 692 621223 74467 74484 CAAGGTAAATTTCTTTCT 69 70430 70447 5-8-5 693 621224 74647 74664 TCAGGCCTGATCTAAGTA 14 70610 70627 5-8-5 694 621225 74827 74844 TCCCCAACCATGTGGTCT 40 70790 70807 5-8-5 695 621226 75007 75024 TGCCAGCCACACCCATGG 52 70970 70987 5-8-5 696 621227 75187 75204 AACTCCACGGCTTCCATG 50 71150 71167 5-8-5 697 621228 75371 75388 AGATAGAAATCTGAAACG 34 71334 71351 5-8-5 698 621229 75551 75568 GCTGGGCCGAGCTGCATT 66 71514 71531 5-8-5 699 621230 75891 75908 GGATTCAAAGGAGAAAAC 35 71854 71871 5-8-5 700 621231 76192 76209 ATTATTATTTGACATGGG 75 72155 72172 5-8-5 701 621232 76373 76390 AGGGTAGGCAGTGTTGTG 4 72336 72353 5-8-5 702 621233 76553 76570 CTCTCTTTGTCAGGAAAA 58 72516 72533 5-8-5 703 621234 76942 76959 CCTTTTTTTTTAGGACGG 0 72905 72922 5-8-5 704 621235 77222 77239 GAGTTGACTGGGCACGGT 33 73185 73202 5-8-5 705 621236 77414 77431 TCATCTGTGAAGCGGACG 81 73377 73394 5-8-5 706 621237 77594 77611 GGCACAATCCATATGAGG 56 73557 73574 5-8-5 707 621238 77781 77798 GTTATTTTGGAACAGTTT 87 73744 73761 5-8-5 708 621239 78117 78134 CTGGACTTTATTTCATTT 77 74080 74097 5-8-5 709 621240 78319 78336 CAGTATATATAGTGCATA 76 74282 74299 5-8-5 710 621241 78499 78516 CAAAACAGCTCCTTGTAA 14 74462 74479 5-8-5 711 621242 78679 78696 ACTTTCATCTACTTTTCA 32 74642 74659 5-8-5 712 621243 78859 78876 CCAATCTGTCCCCAGCTT 42 74822 74839 5-8-5 713 621244 79039 79056 TCCAGCATGAACATAGCT 65 75002 75019 5-8-5 714 621245 79219 79236 TAGAGTCAGTTTCAGGAT 75 75182 75199 5-8-5 715 621246 79399 79416 GGTGGTATCACCAACAGC 35 75362 75379 5-8-5 716 621247 79589 79606 TTAAGGAATGGCTCTGGG 42 75552 75569 5-8-5 717 621248 79769 79786 GAACATGAGACATCTTGA 8 75732 75749 5-8-5 718 621249 79949 79966 AGCTGTGCCCAGTTAAAA 53 75912 75929 5-8-5 719 621250 80129 80146 TCTGGTCATATGAGGAAA 55 76092 76109 5-8-5 720 621251 80309 80326 AGTCATTATCATGTCACC 89 76272 76289 5-8-5 721 621252 80489 80506 GCTGAGCGAATTACCTAA 75 76452 76469 5-8-5 722 621253 80669 80686 CCTCTGTATGACAGAAAT 48 76632 76649 5-8-5 723 621254 80849 80866 GGTAACATGTAAAGCTTC 79 76812 76829 5-8-5 724 621255 81033 81050 TGGATTATGTACAGATAT 28 76996 77013 5-8-5 725 81136 81153 77099 77116 621256 81110 81127 ATGGATGATAGGAAGGAT 20 77073 77090 5-8-5 726 81213 81230 77176 77193 621257 81393 81410 GATGGAGGAAGGAATGAT 5 77356 77373 5-8-5 727 621258 81629 81646 CTGGATGGTTACATGGAT 31 77592 77609 5-8-5 728 621259 81812 81829 ATAATTGATGTATAATTA 0 77775 77792 5-8-5 729 621260 81992 82009 GTTCTGCCCCACCTGGGC 56 77955 77972 5-8-5 730 621261 82198 82215 GCCTCTCAGGGCCTCCGT 76 78161 78178 5-8-5 731 621262 82378 82395 GAAGGATGGCCACACAGA 39 78341 78358 5-8-5 732 621263 82558 82575 ATGGGCTTATCAATGCAT 85 78521 78538 5-8-5 733 621264 82738 82755 GAGGCCCAAATGATCACA 56 78701 78718 5-8-5 734 621265 82918 82935 GCTCAGGGCAGACACGGT 66 78881 78898 5-8-5 735 621266 83098 83115 CTTAACCAGCTAGTGGTG 38 79061 79078 5-8-5 736 621267 83278 83295 GTCCTGTGGAGCTGAAAA 54 79241 79258 5-8-5 737 621268 83486 83503 GGCTGGAGGCATGGAGGG 7 79449 79466 5-8-5 738 621269 83666 83683 TGCCTCCAGAGCACACAC 55 79629 79646 5-8-5 739 621270 83846 83863 AGGATACTAAACCAAGAT 24 79809 79826 5-8-5 740 621271 84026 84043 CACAGATGGGAAGCAAGA 20 79989 80006 5-8-5 741 621272 84206 84223 CATGAGGCAACAATCCAA 70 80169 80186 5-8-5 742

TABLE 20 Inhibition of tau mRNA by 5-10-5 MOE and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 SEQ SEQ ID ID NO: 1 NO: 1 SEQ ISIS Start Stop % ID NO Site Site Sequence inhibition Motif NO 613412 73879 73898 GTGATCTTCCATCACTTCGA 80 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 92 5-8-5 665 98928 98945 621273 84386 84403 GCCGTAAGACCCAGCTCT 58 5-8-5 743 621274 84566 84583 GAAGAAGTACTGAGAAAG 14 5-8-5 744 621275 84746 84763 TAAGAACCTCAGCGGCAA 56 5-8-5 745 621276 84926 84943 TGAGGGAGCACTGAGAGT 21 5-8-5 746 621277 85106 85123 AGTTCAATGAAGGACATT 69 5-8-5 747 621278 85287 85304 ACTGTTGGATGGGTCCAC 70 5-8-5 748 621279 85467 85484 TTGGTCCATTTTGATTGG 0 5-8-5 749 621280 85647 85664 CGGCAGCCCTGTGTAAAC 48 5-8-5 750 621281 85828 85845 CCTTTGCTTCTTGCGCAG 66 5-8-5 751 621282 86014 86031 AGGGCCTTGACTGCCTGG 15 5-8-5 752 621283 86194 86211 AAGGCCCTAAATGCTCTG 57 5-8-5 753 621284 86384 86401 CGCTCAGGTGATCTTGGG 76 5-8-5 754 621285 86564 86581 CTGATTGATTCCCCATCA 64 5-8-5 755 621286 86746 86763 TTCTTGTTGGGCAACTGG 59 5-8-5 756 621287 86928 86945 GAAATGCACTCAGAAGGG 52 5-8-5 757 621288 87108 87125 CCTCAGTGAAATAACTGT 55 5-8-5 758 621289 87288 87305 TGGCTGCAACTTTGAATG 0 5-8-5 759 621290 87470 87487 CTGGAGATGGCAGGCTGG 39 5-8-5 760 621291 87653 87670 CCCGTGGGCCCCATGTGG 1 5-8-5 761 621292 87834 87851 ATTGAGGACACCTGGTGT 6 5-8-5 762 621293 88014 88031 AAGGTCTGCATTGTCAGT 61 5-8-5 763 621294 88194 88211 CTGCTGGCCTCTCTGTAC 56 5-8-5 764 621295 88374 88391 TCGGACACGGTCACTGCC 60 5-8-5 765 621296 88554 88571 GTGCCTTGTCATGTGACA 79 5-8-5 766 621297 88734 88751 GGAAGCCATGTGGTAGCC 50 5-8-5 767 621298 88918 88935 CAGAGGCCACTGCCTCTG 0 5-8-5 768 621299 89098 89115 TTAATAGTACCAAAATCA 25 5-8-5 769 621300 89278 89295 TACATTTCCTTCCTCCCA 10 5-8-5 770 621301 89470 89487 CGGAGTCAGGCAGATGGG 43 5-8-5 771 621302 89650 89667 GCTATGACCTAGTAGGAA 84 5-8-5 772 621303 89830 89847 ATTGTTTACTAGAAACCA 39 5-8-5 773 621304 90010 90027 GGGCAGAGCCCACCCCAT 40 5-8-5 774 621305 90190 90207 AGGCTGGACAAGGCTAGC 63 5-8-5 775 621306 90531 90548 CATATTTGAGTTTCTTTT 62 5-8-5 776 621307 90711 90728 AATGATTACACAAAGCTG 61 5-8-5 777 621308 90891 90908 GAACAGTGTCTTTCCAGC 73 5-8-5 778 621309 91071 91088 ATGTGCCTTTCTGTGCCA 89 5-8-5 779 621310 91251 91268 AATGATTTCTAGAGGTCA 54 5-8-5 780 621311 91431 91448 CCATGATTCCAGGCTGCT 80 5-8-5 781 621312 91816 91833 CAGCCAGGCTAGTCTTGC 80 5-8-5 782 621313 92037 92054 AGGAGAATGTTCTTTTTT 53 5-8-5 783 621314 92219 92236 TTACTGTACTCTGATATA 41 5-8-5 784 621315 92399 92416 CCTTATGGGAGCCCTTTG 54 5-8-5 785 621316 92579 92596 CAGAGCGCAGCAGCACCT 64 5-8-5 786 621317 92759 92776 CTCACCAGCTCTGCTTTA 53 5-8-5 787 621318 92939 92956 CAGCTGATCAAGGGCACA 84 5-8-5 788 621319 93123 93140 AGACCTGCCATAAGACTC 39 5-8-5 789 621320 93380 93397 CTGACTAAAATGTTATTT 34 5-8-5 790 621321 93570 93587 ACAGGCATGAGTTTTTTT 61 5-8-5 791 621322 93840 93857 CCCCTTTTTTTTGGTTTG 2 5-8-5 792 621323 94020 94037 TCTGGAGCCGCCCTGGGC 36 5-8-5 793 621324 94200 94217 ATGACAGCTCTCCTGGTC 38 5-8-5 794 621325 94380 94397 ACTCTCAACTCCTCTGGT 49 5-8-5 795 621326 94576 94593 TCCTGTTGGAGGAGGCGC 7 5-8-5 796 621327 95433 95450 CCTTTGAGTTGAGGGACC 60 5-8-5 797 621328 95613 95630 GCCTGAGCACGGGAGGAG 41 5-8-5 798 621329 95793 95810 GACACCATGAGGGCACCC 69 5-8-5 799 621330 95986 96003 AGCACCTCCTGGGAGGCG 18 5-8-5 800 621331 96166 96183 GGTGGCATGTGGACCAGG 50 5-8-5 801 621332 96346 96363 TGTGGCTCTGAGGTCTCC 72 5-8-5 802 621333 96526 96543 GCCTCTGTCTTAACTTTT 50 5-8-5 803 621334 96778 96795 TTAGGGTACAGTGGTGTG 18 5-8-5 804 621335 96958 96975 CCTCTAGTAGGCCAGTAT 62 5-8-5 805 621336 97141 97158 AATCAAGTAAGTTTGGGA 57 5-8-5 806 621337 97321 97338 CAACTGGGTCTGTAAGCC 47 5-8-5 807 621338 97506 97523 TAGATTTTCTTCTTTGGT 45 5-8-5 808 621339 97686 97703 GGGCACGAATTCTCACTG 53 5-8-5 809 621340 97866 97883 TCCAGAATAGGAGAGGCT 9 5-8-5 810 621341 98046 98063 ACACTGCAGCCAGGCTTG 63 5-8-5 811 621342 98226 98243 TGCAGTCTCCCTAACCCA 59 5-8-5 812 621343 98406 98423 CTCTTTTCCCTTGAATCT 33 5-8-5 813 621344 98607 98624 GTCAGCTTACCTTGGCTT 44 5-8-5 814 621345 98802 98819 CAACTGCTCTTCCCTGGG 68 5-8-5 815 621346 98982 98999 TCTGGGTGCAGTTTATGC 80 5-8-5 816 621347 99162 99179 AGAATTTATGCTGGAAAT 38 5-8-5 817 621348 99342 99359 GGCCAGCTCTCCAAATCC 56 5-8-5 818 621349 99522 99539 CCCTAGACTCTGGGCAAC 66 5-8-5 819

TABLE 21 Inhibition of Tau mRNA by 5-10-5 MOE and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 SEQ SEQ ID ID NO: 1 NO: 1 ISIS Start Stop % SEQ NO Site Site Sequence inhibition Motif ID NO 613412 73879 73898 GTGATCTTCCATCACTTCGA 91 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 96 5-8-5 665 98928 98945 620965 20187 20204 TCTAAGTAATTCGATATG 27 5-8-5 820 620966 20375 20392 AAACAGAAACTAGGGTGG 63 5-8-5 821 620967 20571 20588 TCCCGCTGGGATCCATGG 40 5-8-5 822 620968 20751 20768 TGTTGCCATTGGGCAGCC 0 5-8-5 823 620969 20931 20948 GACCTTCTAATTAGTCCA 58 5-8-5 824 620970 21291 21308 GCGATTCTCTCCTGTCTC 53 5-8-5 825 620971 21567 21584 ACTCAGATTTATTATTTT 87 5-8-5 826 620972 21747 21764 AAAGTACTATCCACTTGT 77 5-8-5 827 620973 21927 21944 CAAATCCTGGGTTCGAAT 39 5-8-5 828 620974 22107 22124 AGGTCTCTGCCCATGAAA 59 5-8-5 829 620975 22287 22304 AGATTTCTCCCTTCACCC 46 5-8-5 830 620976 22467 22484 TTAACATGATGGTGGTTT 86 5-8-5 831 620977 22652 22669 CATTGTGGAAATTAAACA 50 5-8-5 832 620978 22837 22854 TGTAGTCAATATTGAACT 86 5-8-5 833 620979 23269 23286 TTTTGAGAGCAGGTCTCG 61 5-8-5 834 620980 23449 23466 ATGGCAAACAGTACATGT 80 5-8-5 835 620981 23631 23648 AGGATTTAAGTGATTTTC 66 5-8-5 836 620982 23826 23843 TACAAAGACCCCTTTTCC 51 5-8-5 837 620983 24016 24033 AGCTGCCAGCAGTCCTTG 81 5-8-5 838 620984 24196 24213 CTGCTGTAGGAAATGACC 77 5-8-5 839 620985 24377 24394 AGCTCCAACCAGCTTTCT 58 5-8-5 840 620986 24557 24574 GTCCTGATTAACAGCTAA 69 5-8-5 841 620987 24744 24761 GTCTGTCTGCTGCCATCC 81 5-8-5 842 620988 24924 24941 GTAACTGATATGGTAACA 85 5-8-5 843 620989 25286 25303 AGCCCTCTTTGTACAGGA 75 5-8-5 844 620990 25466 25483 CTGTTCCAGTTGATCCAG 53 5-8-5 845 620991 25668 25685 GCAGTCTTTTTAAATTAA 80 5-8-5 846 620992 25848 25865 CCTCTGCTCACATAGAAA 64 5-8-5 847 620993 26028 26045 AGTACCTGCCTTGTTCCT 52 5-8-5 848 620994 26208 26225 AAGCATGGTAATACAAAA 78 5-8-5 849 620995 26404 26421 TTGATTAAAAAAAATAGC 4 5-8-5 850 620996 26584 26601 TTCATCCTTTCCAAGTGT 58 5-8-5 851 620997 26764 26781 CATGCTTACACACCACAC 23 5-8-5 852 26948 26965 620998 26762 26779 TGCTTACACACCACACAC 38 5-8-5 853 26946 26963 620999 27140 27157 ATATGCTGAACACACACA 75 5-8-5 854 621000 27320 27337 CCACCGAGGTCTCATTGG 52 5-8-5 855 621001 27500 27517 GGGAGACCTCCCTTTCAA 0 5-8-5 856 621002 27680 27697 TCTCTGGGTGTAGAGACG 54 5-8-5 857 621003 27861 27878 GCAGGCTCGGGCTCCACG 61 5-8-5 858 621004 28041 28058 CCTTGTCAGATGGTTGAT 55 5-8-5 859 621005 28221 28238 TCTGCAGACTTTTCTCAA 22 5-8-5 860 621006 28426 28443 CTGGAGGGAGGTGATGTG 7 5-8-5 861 621007 28616 28633 GAAAGCTGCTGCTGAGGG 63 5-8-5 862 621008 28981 28998 GGGCAAAGATAATTCGAA 61 5-8-5 863 621009 29165 29182 CCTCACCCTGGGCAAGAA 68 5-8-5 864 621010 29409 29426 TTGCTGCTCGGGAGGCCG 14 5-8-5 865 621011 29589 29606 AAATGAAAGATGCTGGCT 23 5-8-5 866 621012 29953 29970 CAGAGAGGTCCAGCTACT 58 5-8-5 867 621013 30133 30150 AGCTCACTACAGCAGGCA 93 5-8-5 868 621014 30607 30624 TATTTCTTGCAATTCTTT 78 5-8-5 869 621015 30787 30804 GGAGGACTGTCTCTAGAG 59 5-8-5 870 621016 30967 30984 TCCGGCCTGGCGAACTGA 40 5-8-5 871 621017 31278 31295 CCTGCAAAGCTAATTTTA 18 5-8-5 872 621018 31458 31475 GAGCCCCCTTTAAGCCTT 65 5-8-5 873 621019 31654 31671 GTACAAAGACTGTGTACG 47 5-8-5 874 621020 31834 31851 GACTCAGGAACATGTTAG 81 5-8-5 875 621021 32023 32040 TGAGGCAGCAATGCTGGG 34 5-8-5 876 621022 32203 32220 CACTGGGAGGTGTATAGA 0 5-8-5 877 621023 32383 32400 GCGAGGTGGGTGAAGGTT 25 5-8-5 878 621024 32563 32580 AAAAACTTTGGCAGTCAG 80 5-8-5 879 621025 32743 32760 GGTCTGCCCTGCACCAGG 59 5-8-5 880 621026 32923 32940 CTGAGCTCTCCGAGCTGC 61 5-8-5 881 621027 33103 33120 GTGAGGTGGTATCATTGG 51 5-8-5 882 621028 33283 33300 CTAGCTTGAATTCCTCCA 81 5-8-5 883 621029 33463 33480 AATCTGTGACTCAAGAAC 47 5-8-5 884 621030 33708 33725 GGGCCGTCTCTATTAAAA 56 5-8-5 885 621031 33888 33905 GGTATAATTTGTTTGGAC 85 5-8-5 886 621032 34068 34085 CTTTATTGCATATAGGTA 86 5-8-5 887 621033 34248 34265 ATCCAGTCCCAACATTGG 30 5-8-5 888 621034 34428 34445 CGGTCTCTCTGACTTGCC 78 5-8-5 889 621035 34620 34637 GAGGGCATTCCTCAAAGG 10 5-8-5 890 621036 34800 34817 AGTTCTCTGATTCTCAAA 63 5-8-5 891 621037 34980 34997 CCCCCAGTGCCCACAAGT 48 5-8-5 892 621038 35160 35177 CTAAAAGCTAAAGTGGGT 49 5-8-5 893 621039 35340 35357 AGGACCTGGCAGAGCTGC 73 5-8-5 894 621040 35520 35537 AAATCGAACACTTACATA 20 5-8-5 895 621041 35721 35738 TTGCCATCTTGGACAGGG 89 5-8-5 896

TABLE 22 Inhibition of Tau mRNA by 5-10-5 and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 SEQ SEQ ID ID NO: 1 NO: 1 ISIS Start Stop % SEQ ID NO Site Site Sequence inhibition Motif NO 613412 73879 73898 GTGATCTTCCATCACTTCGA 85 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 93 5-8-5 665 98928 98945 620888 98881 98898 ACACCTTCATTTACTGTC 95 5-8-5 897 98918 98935 620889 98890 98907 TTTTCAAACACACCTTCA 74 5-8-5 898 98927 98944 620890 98892 98909 GGTTTTCAAACACACCTT 96 5-8-5 899 98929 98946 620891 98893 98910 TGGTTTTCAAACACACCT 96 5-8-5 900 98930 98947 620892 6191 6208 TACCTGATAGTCGACAGA 53 5-8-5 901 620893 6371 6388 GTCCCTTTCCAGGCCGCC 70 5-8-5 902 620894 6551 6568 CGCCCATTGCGGCAAAAG 73 5-8-5 903 620895 6736 6753 CACTGCTCGGGAGGTGCA 62 5-8-5 904 620896 6938 6955 TGTTCTCAGGCACGGCGC 74 5-8-5 905 620897 7121 7138 GCCCTGCGCTCCGAGCGC 44 5-8-5 906 620898 7301 7318 AAAAAAGATGGCACCTCG 30 5-8-5 907 620899 7481 7498 ACAAAGCAAAGAGCCCCC 59 5-8-5 908 620900 7687 7704 TCACGCCCGTTCCATGCG 43 5-8-5 909 620901 7867 7884 CCGGATGGGTAGCCAGCG 47 5-8-5 910 620902 8047 8064 CCCTCTCCGGACACCTGT 69 5-8-5 911 620903 8227 8244 AAATACACCCAGGGCCGC 51 5-8-5 912 620904 8408 8425 GATAGAATTAACCAGAAA 33 5-8-5 913 620905 8588 8605 AGCCTCGCAGTACCCAGG 39 5-8-5 914 620906 8775 8792 TCGCGAGATGGCAATACG 55 5-8-5 915 620907 8956 8973 TTCCTCCATTAACAGCGC 49 5-8-5 916 620908 9157 9174 CCCGCCTGCTGGGAATGG 65 5-8-5 917 620909 9337 9354 AGAACTCAAATTGGTCCT 67 5-8-5 918 620910 9517 9534 TATAAGCAGCTTATACAG 34 5-8-5 919 620911 9697 9714 CCAAGCCAGGTTATTGCT 70 5-8-5 920 620912 9877 9894 TCTAACAATTTATGGGCA 81 5-8-5 921 620913 10057 10074 AACAAATGGACTGTAACA 59 5-8-5 922 620914 10240 10257 TTCAAGGAGGGAGTAAGG 12 5-8-5 923 620915 10420 10437 AGGAAAGTGGTGGTGGAG 22 5-8-5 924 620916 10600 10617 TGCAGCCCAGCTGGTGAG 35 5-8-5 925 620917 10781 10798 TCCTTGTAGGATGTTTAA 30 5-8-5 926 620918 10961 10978 ACCTGCTACATAGCGGAA 87 5-8-5 927 620919 11141 11158 TTATAGATCATATCTGGG 82 5-8-5 928 620920 11321 11338 AAACAGAGCACCAATGTA 36 5-8-5 929 620921 11501 11518 ACCTCAGAGACTTGACCT 54 5-8-5 930 620922 11681 11698 CCCAGGAAGTGAGAAAAG 20 5-8-5 931 620923 11864 11881 CAGTTGGGAGCCATCTGG 51 5-8-5 932 620924 12044 12061 ATTGGGCTCAGCATGGAG 30 5-8-5 933 620925 12224 12241 TCCTTAACCATTAGGCCA 70 5-8-5 934 620926 12404 12421 TGGCCCTAGTGAGTACCG 75 5-8-5 935 620927 12597 12614 GCCCAGTTTGGTGCAGGG 79 5-8-5 936 620928 12777 12794 GGCGGGAAGGCAGGTCAG 51 5-8-5 937 620929 12957 12974 CAGTTTACCAATGTCAGC 77 5-8-5 938 620930 13137 13154 ATATATTGTTACTTAGTC 83 5-8-5 939 620931 13317 13334 CATAGGCAAAACAGACAA 68 5-8-5 940 620932 13504 13521 TAGTTGAATGTTTTGGAA 73 5-8-5 941 620933 13702 13719 ACGGCGGAGGCTGAGGCG 10 5-8-5 942 620934 13882 13899 AGGTGCAAGCTGGCCGGG 44 5-8-5 943 620935 14062 14079 TGATACCCTGTAAGAATA 82 5-8-5 944 620936 14243 14260 AATATAGCAAAGGGAATT 54 5-8-5 945 620937 14423 14440 TAGAGATAATTATGTCCC 73 5-8-5 946 620938 14609 14626 GCAATTACAGAGCCAGGG 76 5-8-5 947 620939 14789 14806 GCAGAACTGGATCCGATC 69 5-8-5 948 620940 15035 15052 CAAAGCCAGGATGGTCTC 92 5-8-5 949 620941 15246 15263 CCAAAGAGATTCCTTTTT 79 5-8-5 950 620942 15924 15941 GGCCTCATTTATATATAT 20 5-8-5 951 620943 16156 16173 TATGGGACAATCATAGCT 56 5-8-5 952 620944 16336 16353 AATTCTGATACATTGTCA 81 5-8-5 953 620945 16516 16533 GATGACTCTTCCAAATGG 70 5-8-5 954 620946 16696 16713 TCACTTCATTATTCAAGC 84 5-8-5 955 620947 16876 16893 GTGCAGCCGCCACTGGCC 86 5-8-5 956 620948 17085 17102 CGATAAGATCTTTTTAAA 25 5-8-5 957 620949 17266 17283 GACAGTATAATCTCCATT 69 5-8-5 958 620950 17446 17463 AGGCTAAAGTCAGCCCAC 29 5-8-5 959 620951 17639 17656 CTTGCACCCTGGTTTGGG 40 5-8-5 960 620952 17819 17836 TCCCTGTCTGGTTAGGAA 80 5-8-5 961 620953 17999 18016 ACGCGCCTGCCCCGTGCC 50 5-8-5 962 620954 18179 18196 CTCAAACCTGGCCACGCT 64 5-8-5 963 620955 18359 18376 GAAACTGGCGGGTCTGTT 64 5-8-5 964 620956 18539 18556 GGGAGGAATTTGGCAATG 37 5-8-5 965 620957 18719 18736 AAAGAGCACGGCATCTAT 76 5-8-5 966 620958 18905 18922 AAACTGTGAGGCACTGGG 87 5-8-5 967 620959 19088 19105 CATTTGACATTGGCCTGT 72 5-8-5 968 620960 19283 19300 ATTATTATCAGCATCTTC 77 5-8-5 969 620961 19467 19484 TTGCAGAGGCAGGAAGGC 67 5-8-5 970 620962 19647 19664 AGCAGCCTGGTTTAGAGG 31 5-8-5 971 620963 19827 19844 TCCACATCTTCCAGCCTC 74 5-8-5 972 620964 20007 20024 ATACAGACTCAGTCTCCT 79 5-8-5 973

TABLE 23 Inhibition of Tau mRNA by 5-10-5 and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 SEQ SEQ ID ID NO: 1 NO: 1 ISIS Start Stop % SEQ NO Site Site Sequence inhibition Motif ID NO 613412 73879 73898 GTGATCTTCCATCACTTCGA 78 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 93 5-8-5 665 98928 98945 621042 35901 35918 AGTCGACCGCACTCCTGC 42 5-8-5 974 621043 36106 36123 GCCCTGAGGGTCCACAGG 20 5-8-5 975 621044 36286 36303 AGGGCAGTGTCCTACCTC 50 5-8-5 976 621045 36466 36483 GTCCTCCCTTCCCCACAA 29 5-8-5 977 621046 36653 36670 CTGAATTCCTATCATGCG 51 5-8-5 978 621047 36833 36850 GAGAAAGAAGAGTGTGGT 23 5-8-5 979 621048 37024 37041 CTTAGGACAGATTCCTAG 29 5-8-5 980 621049 37204 37221 GTCAGAAGTGTTTTCCCA 80 5-8-5 981 621050 37413 37430 TCCAATTTTAAACTTAAA 28 5-8-5 982 621051 37750 37767 CTACTGTTTAAAAAGGCT 66 5-8-5 983 621052 38013 38030 GTAAACACCATCTCTAAA 19 5-8-5 984 621053 38193 38210 CTCTGATACTATTATAAG 31 5-8-5 985 621054 38374 38391 CTGAGCTATTTGAGAATT 21 5-8-5 986 621055 38557 38574 TCTGCATATATAATTATA 57 5-8-5 987 621056 38737 38754 CACATTGTTTAGATTTTC 79 5-8-5 988 621057 38922 38939 TTAGCACACCTGAAACGC 51 5-8-5 989 621058 39102 39119 GGCAAAGAAGACAGGAGA 68 5-8-5 990 621059 39541 39558 TGCTGAGGATGCTTTTTT 72 5-8-5 991 621060 39721 39738 GCTGGAAGATGAGGTGGT 47 5-8-5 992 621061 39901 39918 TGGGTGTTGGAGAAAACT 24 5-8-5 993 621062 40100 40117 GCTTTAAAAGCTCCTCAG 65 5-8-5 994 621063 40347 40364 CGAGCACTTGTAGTTCCA 35 5-8-5 995 621064 40528 40545 GCAGTGCTTTAAAAATAT 22 5-8-5 996 621065 40709 40726 GAAATTTCATGAAATATG 37 5-8-5 997 621066 40889 40906 TTACTTGCAATTGAAAGA 6 5-8-5 998 621067 41069 41086 GGAAAGACTACTCAGAGC 57 5-8-5 999 621068 41249 41266 GATGAAAGAGAATAACAA 20 5-8-5 1000 621069 41429 41446 ACCCATTATCTGCTCCCC 66 5-8-5 1001 621070 41611 41628 TTTTCAGTAACAACATAA 51 5-8-5 1002 621071 41944 41961 CCTTTAGCTTTCCTTTTC 47 5-8-5 1003 621072 42124 42141 CAAATTCTGTTTCTGTAA 47 5-8-5 1004 621073 42304 42321 GCAAAAATAAGTGAACTG 38 5-8-5 1005 621074 42492 42509 TTGCAGTGACTTCTTGGG 71 5-8-5 1006 621075 42672 42689 AAACCCCTTCAACATCAG 45 5-8-5 1007 621076 42853 42870 ACAAGAGAAACATTTTAC 53 5-8-5 1008 621077 43033 43050 CACTTAGTAACCAAGCAA 53 5-8-5 1009 621078 43213 43230 CATAGATGATATATTTTG 78 5-8-5 1010 621079 43393 43410 AAATCTTGAACTCCTGAA 58 5-8-5 1011 621080 43620 43637 GGAGTTTTTGTGTTTCTT 77 5-8-5 1012 621081 43804 43821 AAATTCTTAATGGTTCAG 75 5-8-5 1013 621082 44051 44068 AGATAGATCTCGGCTCAC 81 5-8-5 1014 621083 44231 44248 AACCTTTAATAAACTTTT 43 5-8-5 1015 621084 44617 44634 GTCCAAGAGGTTTTTTTT 41 5-8-5 1016 621085 44809 44826 GCAGTCACTGCATTCCAG 27 5-8-5 1017 621086 45050 45067 CAAAAACAGGGCTAGGCA 21 5-8-5 1018 621087 45230 45247 TTCCCTGAACAAATGGCA 59 5-8-5 1019 621088 45412 45429 CTCAAATGTAACATTTTA 84 5-8-5 1020 621089 45592 45609 TACACCTGGCCCATGACC 41 5-8-5 1021 621090 45772 45789 AAGTTCTGGCTCAAGCAA 21 5-8-5 1022 621091 45955 45972 ACAACTCTCTGGAAAAAA 30 5-8-5 1023 621092 46135 46152 AGTCCACGCCTGCATCTG 63 5-8-5 1024 621093 46315 46332 CAAGGCAACAGAGAGGCA 42 5-8-5 1025 621094 46495 46512 AACATGCATATAATTACC 40 5-8-5 1026 621095 46675 46692 TTGGAGGGACAATTCTCA 46 5-8-5 1027 621096 46855 46872 CGAAGAGGCCAGGACTGC 34 5-8-5 1028 621097 47036 47053 TTTGGCTCTGGTGATGGT 45 5-8-5 1029 621098 47216 47233 ACCAGGCAAAGGGACACC 68 5-8-5 1030 621099 47396 47413 TGCCAGAGACAGAGGTTG 68 5-8-5 1031 621100 47576 47593 CACACGCATGGCTTCATG 55 5-8-5 1032 621101 47756 47773 GAGGAAGGCATGAACTAA 60 5-8-5 1033 621102 47936 47953 TCCCATGCCCACGCATGT 26 5-8-5 1034 621103 48116 48133 ATTATTAACGAACAAAAA 2 5-8-5 1035 621104 48302 48319 ATGTCCAGATGTGGATCG 52 5-8-5 1036 621105 48482 48499 ATCAGACAGAAGAGCCAT 55 5-8-5 1037 621106 48678 48695 GCACAGTGCAACAGTGGG 78 5-8-5 1038 621107 49171 49188 GGCTTTCCCTTCCCTTCT 33 5-8-5 1039 621108 49361 49378 AGCTGGGAAAGTGGCAGG 23 5-8-5 1040 621109 49561 49578 ATGACAACTTGATTTGGG 65 5-8-5 1041 621110 49741 49758 GACCGCAACCTTGCCAAA 43 5-8-5 1042 621111 50111 50128 CTTTCAGAGACAGGCTCG 41 5-8-5 1043 621112 50298 50315 GGGCTCACCCCTGTAGTT 9 5-8-5 1044 621113 50766 50783 CTCTTAAGGCCGAGTGCA 39 5-8-5 1045 621114 50970 50987 TGCTAGGCAAAAAAACAA 35 5-8-5 1046 621115 51150 51167 TAATTAGAAACCTCCTCG 10 5-8-5 1047 621116 51330 51347 CAGGTCATATATGAAATC 33 5-8-5 1048 621117 51521 51538 TGTGCTTCCGAAAGTAGT 63 5-8-5 1049 621118 51702 51719 GCATCAAATCCATGCATT 38 5-8-5 1050

TABLE 24 Inhibition of tau mRNA by 5-10-5 and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 SEQ SEQ ID ID NO: 1 NO: 1 ISIS Start Stop % SEQ NO Site Site Sequence inhibition Motif ID NO 613412 73879 73898 GTGATCTTCCATCACTTCGA 71 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 84 5-8-5 665 98928 98945 621119 51894 51911 ACTCTGAAGCACAGACAG 21 5-8-5 1051 621120 52081 52098 CTTACCTGGGCTGGATGG 5 5-8-5 1052 621121 52261 52278 TTCTGTCTATAGCTAGAA 32 5-8-5 1053 621122 52441 52458 AGATGGCTTCCTATTGTT 32 5-8-5 1054 621123 52621 52638 TGGTTTGGTCAGGCCTCA 51 5-8-5 1055 621124 52803 52820 ATTGTAACTCAAAGTGGG 50 5-8-5 1056 621125 53004 53021 AAACTCTACACCCCTGGG 17 5-8-5 1057 621126 53264 53281 CATACCAGGAAGGAAGGA 17 5-8-5 1058 621127 53444 53461 TGGACATATCAGAATTTG 64 5-8-5 1059 621128 53624 53641 CCCTTATTAGTGTCTACA 68 5-8-5 1060 621129 53805 53822 ACACTCTAGAGAGGTCCA 68 5-8-5 1061 621130 53985 54002 GACAGTACCCACGACACG 32 5-8-5 1062 621131 54165 54182 CCCCATTCTGGATCCTGG 40 5-8-5 1063 621132 54348 54365 ATTGTCTGTAAAATGTGG 57 5-8-5 1064 621133 54528 54545 ATTCTGTCTGACAGAGAC 40 5-8-5 1065 621134 54711 54728 CGATGGTGCACATCTATA 45 5-8-5 1066 621135 54891 54908 CAGTGTCCGCTGAACACA 37 5-8-5 1067 621136 55073 55090 AGGAGGATTAGGAAGAAA 14 5-8-5 1068 621137 55261 55278 CAAGGAAGGCGATCTGGG 17 5-8-5 1069 621138 55623 55640 GGTGGCACTTGAACAAGC 61 5-8-5 1070 621139 55803 55820 GGTGGAAAGGGAACCCGG 43 5-8-5 1071 621140 55983 56000 CCCCACGGGTACACAGAG 20 5-8-5 1072 621141 56163 56180 CGCTGCGAACGATGCACT 63 5-8-5 1073 621142 56343 56360 GAATGGTCCTTCTTACAG 13 5-8-5 1074 621143 56526 56543 CTGGGAAAACCAAGGCAA 57 5-8-5 1075 621144 56706 56723 CCCAGGCTGAGGAAGGAC 40 5-8-5 1076 621145 56886 56903 GATTGGCTTTTACATCTT 48 5-8-5 1077 621146 57289 57306 CGGTTTGAGTACAGTGGT 50 5-8-5 1078 621147 57470 57487 CTAACATCTTAGGGCACA 73 5-8-5 1079 621148 57650 57667 TTCCTCTGATGTCAGAAT 51 5-8-5 1080 621149 58090 58107 CCCCCTCGGTCGCCCAGG 37 5-8-5 1081 621150 58271 58288 TGTTTTAACTAAAAGCTT 24 5-8-5 1082 621151 58451 58468 GTATGAGGGCATAGATGA 26 5-8-5 1083 621152 58631 58648 ATCACACCACAGAATGTT 29 5-8-5 1084 621153 58812 58829 TACTCTTCTGTGGCAGCT 66 5-8-5 1085 621154 59198 59215 CAACAGCAAGGCCAGGCG 44 5-8-5 1086 621155 59380 59397 TAGATAATTTGAAATTTA 0 5-8-5 1087 621156 59786 59803 ACCTTAATCCCAGTATTT 12 5-8-5 1088 621157 59966 59983 TGCTTATGGTCCAGCTGT 40 5-8-5 1089 621158 60146 60163 GAGCCAGGCTTCAAAACC 26 5-8-5 1090 621159 60326 60343 TCCAGCACATTTAGGCGG 32 5-8-5 1091 621160 60522 60539 ATCTCAATAAAGCTATCG 70 5-8-5 1092 621161 60702 60719 CGCCCGCGGTGACTCAGC 32 5-8-5 1093 621162 60906 60923 GGAGTCAAGAGTCCAGGG 44 5-8-5 1094 621163 61109 61126 AGAAGGAATGATGAGGAA 24 5-8-5 1095 621164 61290 61307 GCAGTACAGTTTGGTAAT 32 5-8-5 1096 621165 61476 61493 TGGCTACTCTCTCAGGAG 9 5-8-5 1097 621166 61656 61673 GACCCTAGGTGGGTAAAG 21 5-8-5 1098 621167 61836 61853 GTCCCCTGAGGTGTTGTG 0 5-8-5 1099 621168 62016 62033 CCCAGACAACCTTCACCT 34 5-8-5 1100 621169 62208 62225 TGGGTTAAGACTCTGTCT 11 5-8-5 1101 621170 62467 62484 TGGAGCCGAGTGCGGTGG 12 5-8-5 1102 621171 62657 62674 TCTCTTCTGGATATTTCG 49 5-8-5 1103 621172 62843 62860 CGATTTAAAATGAATGAT 20 5-8-5 1104 621173 63023 63040 AGAACAAAGATAACAGTT 0 5-8-5 1105 621174 63218 63235 GCTAAATTGAGGCCCGGG 42 5-8-5 1106 621175 63398 63415 CTTTCTCCTTTCCTCAAT 26 5-8-5 1107 621176 63578 63595 TCTAAGCGAGACACACAG 26 5-8-5 1108 621177 63758 63775 TGATTTCTCTGGAAGAAC 27 5-8-5 1109 621178 63938 63955 GCCTGTGTCTTCTAACTC 32 5-8-5 1110 621179 64184 64201 GGCTGAGAAGCTGGGACC 40 5-8-5 1111 621180 64365 64382 TTTTTACTTGAGATGTCA 38 5-8-5 1112 621181 64543 64560 GTAGGTGTCATCATCATC 83 5-8-5 1113 621182 64723 64740 CAACACTGACTGAGCACA 56 5-8-5 1114 621183 64903 64920 GCATTGGCTGGGCTAGGT 73 5-8-5 1115 621184 65092 65109 TATTTACTATGCAAAATA 18 5-8-5 1116 621185 65273 65290 AGCACCACAGAGCAGATG 19 5-8-5 1117 621186 65453 65470 CTCAGTGTGGAGTTCTGC 44 5-8-5 1118 621187 65634 65651 CATTTTTCAACCACTTAA 44 5-8-5 1119 621188 65815 65832 GTAAGGCTTTGTGGGCCA 32 5-8-5 1120 621189 66148 66165 TACTTTGCATTATTTATT 44 5-8-5 1121 621190 66535 66552 TTGGCTTTCATTATAATT 13 5-8-5 1122 621191 66736 66753 GGACGGTTGGGAAATAGG 18 5-8-5 1123 621192 66916 66933 GAGCAACTGTTCATAGGG 49 5-8-5 1124 621193 67096 67113 TCTGCACATCGACACATC 34 5-8-5 1125 621194 67538 67555 AGGCTAGGCCCCATGGCT 21 5-8-5 1126 621195 67718 67735 GCACCCCCATCCTTCAGC 30 5-8-5 1127

TABLE 25 Inhibition of Tau mRNA by 5-10-5 and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 SEQ ID SEQ ID ISIS NO: 1 start NO: 1 stop % SEQ ID NO site site Sequence inhibition Motif NO: 613412 73879 73898 GTGATCTTCCATCACTTCGA 87 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 96 5-8-5 665 98928 98945 621350 99702 99719 CCAGGAGATGAAGTAGCA 75 5-8-5 1128 621351 n/a n/a ACTCTTGGGAAACAAAGT 31 5-8-5 1129 621352 100065 100082 GGAAGCAGCCAGGGACGG 38 5-8-5 1130 621353 100288 100305 CTGCCTGGCCAATTAAAT 28 5-8-5 1131 621354 100468 100485 CAAACTAATATTAACACT 0 5-8-5 1132 621355 100648 100665 TACTCAATTCCAGGCAAG 71 5-8-5 1133 621356 100836 100853 AATGGAAGTTCTAGTACG 57 5-8-5 1134 621357 101188 101205 CAATCACAGTTCTTTTTC 45 5-8-5 1135 621358 101381 101398 GTCTTAAACATAAACATA 77 5-8-5 1136 621359 101759 101776 TGAGAGGGATGGCCCCCA 67 5-8-5 1137 621360 n/a n/a CGCATAACACCACGCCCG 90 5-8-5 1138 621361 102169 102186 GCCAAAATCAGGAATGGG 94 5-8-5 1139 621362 102349 102366 GGCAGTCCCTGGGTTCCG 77 5-8-5 1140 621363 102529 102546 GTTGAACCTGACCAAGGA 89 5-8-5 1141 621364 102709 102726 AGCATGAGTTGTGCCAAG 89 5-8-5 1142 621365 102889 102906 CCCACTCCGCCACCTTGA 67 5-8-5 1143 621366 103091 103108 TACCAGAGCTGGGTGGTG 13 5-8-5 1144 621367 103271 103288 ACATTTGCCTCAGAAATC 5 5-8-5 1145 621368 103593 103610 GATTCAGGCTGGGAGTGG 11 5-8-5 1146 621369 103773 103790 CTCCGAGAGCTGCCACTT 51 5-8-5 1147 621370 103966 103983 TGCAGCAGACTCCTAACG 33 5-8-5 1148 621371 104389 104406 AGGAATCTCACTTTTGTC 25 5-8-5 1149 621372 104569 104586 AGGCTGCCCGTGCCACCA 30 5-8-5 1150 621373 104749 104766 TTAGTGAATGGCCATCCT 33 5-8-5 1151 621374 105023 105040 AAACTGCTTGAATTTGGG 31 5-8-5 1152 621375 105203 105220 TGTGAGTCCCCTGTAATC 18 5-8-5 1153 621376 105383 105400 TAAACAGGGCCTTAATGA 0 5-8-5 1154 621377 105563 105580 TCCTCCCGGCTCTAGAAA 10 5-8-5 1155 621378 105745 105762 GGAGACAGCCAGGCATGG 18 5-8-5 1156 621379 106173 106190 ATAATGTTTTTTATAGAG 0 5-8-5 1157 621380 106365 106382 GACTGGATTTTTGTATTT 30 5-8-5 1158 621381 106545 106562 CGGAGTCTTATTCTGATG 39 5-8-5 1159 621382 106725 106742 CCGCAGCAATGCCCCTGC 65 5-8-5 1160 621383 106905 106922 TCTCGATCCCCTTCAAGA 23 5-8-5 1161 621384 107085 107102 CTGGGTGTCCTTTACCCT 5 5-8-5 1162 621385 107265 107282 CCTGCTCCACGCCTGCCT 82 5-8-5 1163 621386 107445 107462 TCAGGGCCTTTATCCTAA 51 5-8-5 1164 621387 107625 107642 TCGGCTCCACAGTCTTTG 60 5-8-5 1165 621388 107805 107822 CACCACTGGGTTAGGCAG 38 5-8-5 1166 621389 108170 108187 TCACCTTCCCGCCTCCCG 16 5-8-5 1167 621390 108359 108376 TTGAAGAGGGTCCAGAGG 3 5-8-5 1168 621391 108548 108565 TGCCCAGAAGGCAGGTGG 38 5-8-5 1169 621392 108728 108745 GGCCCCCGAAGTCTGTGC 62 5-8-5 1170 621393 108908 108925 CGGTGCAGGACAGAGGTG 57 5-8-5 1171 621394 109135 109152 CGCGCACCACCACCACGC 78 5-8-5 1172 621395 109315 109332 TCAGGCTGGGCCCTAAGC 50 5-8-5 1173 621396 109495 109512 TCACCCTTCCCCAGCTCC 45 5-8-5 1174 621397 109675 109692 TCAGACCCAAGTGATAAG 64 5-8-5 1175 621398 109855 109872 GCAGGGTCTACACATGCG 76 5-8-5 1176 621399 110045 110062 CTGAGATGTTCTCTTCCT 73 5-8-5 1177 621400 110225 110242 AGCTGCGGTACAGGACAG 68 5-8-5 1178 621401 110405 110422 CCAGTGAGGGCCCCTCTG 20 5-8-5 1179 621402 110585 110602 GCCCCCAGAGGTGCATGG 58 5-8-5 1180 621403 110769 110786 GGAGCAGCAGACACATGC 79 5-8-5 1181 621404 110949 110966 GGAAGCTACTTCCCATGC 43 5-8-5 1182 621406 111495 111512 GGAGGAGGCCTGACACCC 58 5-8-5 1183 621407 111675 111692 TTTCCCTTTGGTGTTAGC 91 5-8-5 1184 621408 111856 111873 CCCCAAAGCAATCTATGT 47 5-8-5 1185 621409 112036 112053 TCTGGAGGGAACACTGCC 60 5-8-5 1186 621410 112216 112233 GGTTGAGGTTGAGGGTAG 37 5-8-5 1187 621411 112396 112413 ACTCCCGCAGGCCAAACA 36 5-8-5 1188 621412 112603 112620 GAGTGCCAACAGGCCCAG 75 5-8-5 1189 621413 112784 112801 TTCTAAGTACAATTTGGG 67 5-8-5 1190 621414 112984 113001 TTACTGGTTGTGTTTTCT 94 5-8-5 1191 621415 113164 113181 GAGTCTCAGTCTCACTGT 76 5-8-5 1192 621416 113347 113364 CATTCCTCCTGCTGCTGT 83 5-8-5 1193 621417 113540 113557 GCTCTGAAGAGCTCCACG 83 5-8-5 1194 621418 113720 113737 CTCTCTTCAGGGCCACCG 82 5-8-5 1195 621419 113900 113917 CTCTCTCGCCCTGCATGG 48 5-8-5 1196 621420 114080 114097 CTGGACATCCTCCGAGAA 27 5-8-5 1197 621421 114260 114277 CACCCAGGCTGCGGCCCA 64 5-8-5 1198 621422 114440 114457 CTGTGGTGCAAGCCTGTG 42 5-8-5 1199 621423 114639 114656 AGACGAGAGTGCGCCCAC 78 5-8-5 1200 621424 115155 115172 CCACATAAATGTTCTACA 94 5-8-5 1201 621425 115335 115352 GGTACAGGAAAAGATGCC 88 5-8-5 1202 621426 115515 115532 ATCAGCTTAGGAACTGAC 85 5-8-5 1203

TABLE 26 Inhibition of tau mRNA by 5-10-5 and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 SEQ ID SEQ ID ISIS NO: 1 start NO: 1 stop % SEQ ID NO site site Sequence inhibition Motif NO: 613412 73879 73898 GTGATCTTCCATCACTTCGA 86 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 97 5-8-5 665 98928 98945 621427 115695 115712 GTGCCTCACAGTTGGCTC 39 5-8-5 1204 621428 115896 115913 GAGAGGCTGTGGTGAGGT 25 5-8-5 1205 621429 116077 116094 GAAATCAGAATCCTGAAA 66 5-8-5 1206 621430 116259 116276 ATGTGGCTTTTTCTTGTT 64 5-8-5 1207 621431 116595 116612 GTTTTTTTCTGGCCGGGC 90 5-8-5 1208 621432 116955 116972 TTTTCAGTGGATAAGGCT 78 5-8-5 1209 621433 117135 117152 ACATAAACTCCTTCTTTG 53 5-8-5 1210 621434 117315 117332 TGGTGAATGTTGAAATTC 78 5-8-5 1211 621435 117495 117512 AGGACAAGGAGGCCATGT 56 5-8-5 1212 621436 117787 117804 TCAAATGACAGATTCTCA 78 5-8-5 1213 621437 117967 117984 CATCCCTACTTTCTCCCT 24 5-8-5 1214 621438 118420 118437 TAGAATCTTGCCTTGTCG 34 5-8-5 1215 621439 118600 118617 GATACAGACATACATTGT 68 5-8-5 1216 621440 118831 118848 AATTCTTTGTGATGATGG 82 5-8-5 1217 621441 119044 119061 GCACATATTTACATTTTA 94 5-8-5 1218 621442 119224 119241 GGTGAGAGAGCCAGCCTC 59 5-8-5 1219 621443 119404 119421 CAAACAGCCTCCCATGAA 38 5-8-5 1220 621444 119584 119601 ATATCCTAAGCATTGTCT 71 5-8-5 1221 621445 119880 119897 TACAGTGGCTCCTGTAAT 18 5-8-5 1222 621446 120060 120077 GCTCATCAAAGCAAAAAC 84 5-8-5 1223 621447 120248 120265 CTGGGCTGTCGGATCTGG 69 5-8-5 1224 621448 120439 120456 AAGCCACCATGCCTGTAA 89 5-8-5 1225 621449 120660 120677 ACACCATCACGGCTCAGT 56 5-8-5 1226 621450 120840 120857 AGGGAGTTGGAAAAACTG 46 5-8-5 1227 621451 121209 121226 CAGGTTTGAGAAGCCCTG 0 5-8-5 1228 621452 121389 121406 AGACCCACCGGCACATTC 56 5-8-5 1229 621453 121569 121586 ATTTCCAGCGGGCTTTAC 62 5-8-5 1230 621454 121761 121778 GCTTGCTCGCAAGGACGC 93 5-8-5 1231 621455 121941 121958 CCACAGCACGGCGCATGG 65 5-8-5 1232 621456 122121 122138 ACACCCCCTCCTAGAATA 0 5-8-5 1233 621457 122301 122318 ACCACCTTCAGCCCAACT 22 5-8-5 1234 621458 122481 122498 TAAGCTGGAGGCTTAGGA 40 5-8-5 1235 621459 122678 122695 GAACCTCCCGCCTTAGTC 19 5-8-5 1236 621460 122913 122930 TATATGATTGTTTTTTGA 0 5-8-5 1237 621461 123093 123110 AGGAGCCTCGGGTGCCCA 30 5-8-5 1238 621462 123273 123290 TCAGGCCAGGCATTTTCT 58 5-8-5 1239 621463 123453 123470 CACTTAGCAGACACTGGT 77 5-8-5 1240 621464 123633 123650 TCCTCTGGCCGAGCTCAC 58 5-8-5 1241 621465 123824 123841 ACAGCGCGGGACACACGG 77 5-8-5 1242 621466 124004 124021 GCATCTCTTCTCACCAGG 13 5-8-5 1243 621467 124184 124201 GGATACCTGGAGGGCAGG 13 5-8-5 1244 621468 124379 124396 TGAGGCCAGCACTGAGGG 29 5-8-5 1245 621469 124559 124576 ACTGGAACCATCCCGAAT 37 5-8-5 1246 621470 124752 124769 GCAGTGGCCTTGTGTGGG 19 5-8-5 1247 621471 124932 124949 ACCCTGAGCTGCCAGCTG 53 5-8-5 1248 621472 125112 125129 CTGCACACTCAGTGTCCT 78 5-8-5 1249 621473 125292 125309 CTGGAAAGGCAGGAGTGG 33 5-8-5 1250 621474 125472 125489 AAGAGCCTCTGGGAAAAA 50 5-8-5 1251 621475 125652 125669 GGTGTAACTCAATGAGAA 41 5-8-5 1252 621476 125832 125849 GGCTACCTGGTTTATGAT 32 5-8-5 1253 621477 126012 126029 GGAGCGAGCTGGAGCCAC 75 5-8-5 1254 621478 126332 126349 GCTCACTCCGCTCACTGC 74 5-8-5 1255 621479 126527 126544 CTAGGTCTTATTCTATTT 38 5-8-5 1256 621480 127208 127225 AGGTTTTTGTTGTGTGTT 54 5-8-5 1257 621481 127396 127413 CCTCTTCACCTTAAAAAA 24 5-8-5 1258 621482 127591 127608 AAACAAAGTCACAGAGGG 67 5-8-5 1259 621483 127934 127951 AGGATCACTTTCTTTTCT 31 5-8-5 1260 621484 128271 128288 TTGCTGAGTCTCACTCTG 87 5-8-5 1261 621485 128451 128468 CAGAATCTATAGCTGTGT 51 5-8-5 1262 621486 128631 128648 ATACTTTCCACAGGGAGA 45 5-8-5 1263 621487 128811 128828 GAAGAAGGGTCCCTCTCT 38 5-8-5 1264 621488 128993 129010 ATGAGAGTACAACTCCAT 44 5-8-5 1265 621489 129269 129286 ACCCTTTCAGGCCGGGCG 44 5-8-5 1266 621490 129678 129695 TAAGTAAAGTCTGTTTTT 47 5-8-5 1267 621491 129861 129878 TAGAATGGTGTTTGGATA 42 5-8-5 1268 621492 130040 130057 GCAGTCTACAGGGCAAAG 79 5-8-5 1269 621493 130234 130251 ACCTTTTTATTTCCTCCG 14 5-8-5 1270 621494 130414 130431 AGCTCCAGGTGATTGAGA 68 5-8-5 1271 621495 130691 130708 GTCGCCATGTAAGAAATG 46 5-8-5 1272 621496 130902 130919 AGGGAGACTGATATGGTT 31 5-8-5 1273 621497 131113 131130 ATATTGCTTGACCTCAGG 18 5-8-5 1274 621498 131543 131560 AGTGGTTCTCGCTTTTTT 75 5-8-5 1275 621499 131725 131742 CAAGACAGGCAGGCAGAG 19 5-8-5 1276 621500 131905 131922 GTGCTTGGCTGCGCTTCT 45 5-8-5 1277 621501 132085 132102 CAGACCTCTATCTTTTAG 24 5-8-5 1278 621502 132266 132283 AGGAGGGTCTTTCCCGTG 57 5-8-5 1279 621503 132446 132463 AACTGGGCCACCATGAGA 30 5-8-5 1280

TABLE 27 Inhibition of Tau mRNA by 5-8-5 MOE gapmers targeting SEQ ID NO: 1 SEQ ID SEQ ID ISIS NO: 1 start NO: 1 stop % SEQ ID NO site site Sequence inhibition Motif NO: 621519 5901 5918 CCTGCCGCTCGGCCGTCC 19 5-8-5 1281 621520 5904 5921 CGCCCTGCCGCTCGGCCG 0 5-8-5 1282 621521 5919 5936 GTGGGCGCGCGCGAGCGC 7 5-8-5 1283 621522 5922 5939 CTAGTGGGCGCGCGCGAG 6 5-8-5 1284 621523 5925 5942 CCACTAGTGGGCGCGCGC 0 5-8-5 1285 621524 5928 5945 CGGCCACTAGTGGGCGCG 18 5-8-5 1286 621525 5931 5948 CTCCGGCCACTAGTGGGC 39 5-8-5 1287 621526 5934 5951 CTCCTCCGGCCACTAGTG 37 5-8-5 1288 621527 5937 5954 CTTCTCCTCCGGCCACTA 15 5-8-5 1289 621528 5940 5957 AGCCTTCTCCTCCGGCCA 0 5-8-5 1290 621529 5943 5960 GGGAGCCTTCTCCTCCGG 0 5-8-5 1291 621530 5946 5963 CGCGGGAGCCTTCTCCTC 28 5-8-5 1292 621531 5949 5966 CTCCGCGGGAGCCTTCTC 0 5-8-5 1293 621532 5952 5969 GGCCTCCGCGGGAGCCTT 0 5-8-5 1294 621533 5955 5972 CGCGGCCTCCGCGGGAGC 0 5-8-5 1295 621534 5958 5975 CAGCGCGGCCTCCGCGGG 0 5-8-5 1296 621535 5961 5978 GGGCAGCGCGGCCTCCGC 31 5-8-5 1297 621536 5964 5981 GGCGGGCAGCGCGGCCTC 17 5-8-5 1298 621537 5987 6004 ACGCGAGCCTCCCCAGGG 10 5-8-5 1299 621538 5990 6007 GGAACGCGAGCCTCCCCA 33 5-8-5 1300 621539 5993 6010 GCGGGAACGCGAGCCTCC 0 5-8-5 1301 621540 5996 6013 GCAGCGGGAACGCGAGCC 0 5-8-5 1302 621541 5999 6016 CGAGCAGCGGGAACGCGA 0 5-8-5 1303 621542 6002 6019 GCGCGAGCAGCGGGAACG 9 5-8-5 1304 621543 6005 6022 CAGGCGCGAGCAGCGGGA 22 5-8-5 1305 621544 6008 6025 GCGCAGGCGCGAGCAGCG 1 5-8-5 1306 621545 6011 6028 GCGGCGCAGGCGCGAGCA 11 5-8-5 1307 621546 6014 6031 CGGGCGGCGCAGGCGCGA 47 5-8-5 1308 621547 6017 6034 CGGCGGGCGGCGCAGGCG 15 5-8-5 1309 621548 6020 6037 GGCCGGCGGGCGGCGCAG 0 5-8-5 1310 621549 6023 6040 TGAGGCCGGCGGGCGGCG 5 5-8-5 1311 621550 6026 6043 TCCTGAGGCCGGCGGGCG 20 5-8-5 1312 621551 6029 6046 CGTTCCTGAGGCCGGCGG 13 5-8-5 1313 621552 6045 6062 GCCGGCGAAGAGGGCGCG 0 5-8-5 1314 621553 6048 6065 CGCGCCGGCGAAGAGGGC 0 5-8-5 1315 621554 6051 6068 GCGCGCGCCGGCGAAGAG 0 5-8-5 1316 621555 6054 6071 AGGGCGCGCGCCGGCGAA 32 5-8-5 1317 621556 6057 6074 GCGAGGGCGCGCGCCGGC 15 5-8-5 1318 621557 6060 6077 ACTGCGAGGGCGCGCGCC 30 5-8-5 1319 621558 6063 6080 GTGACTGCGAGGGCGCGC 38 5-8-5 1320 621559 6066 6083 GCGGTGACTGCGAGGGCG 30 5-8-5 1321 621560 6069 6086 GTGGCGGTGACTGCGAGG 37 5-8-5 1322 621561 6072 6089 TGGGTGGCGGTGACTGCG 48 5-8-5 1323 621562 6075 6092 TGGTGGGTGGCGGTGACT 34 5-8-5 1324 621563 6078 6095 AGCTGGTGGGTGGCGGTG 54 5-8-5 1325 621564 6081 6098 CGGAGCTGGTGGGTGGCG 36 5-8-5 1326 621565 6084 6101 TGCCGGAGCTGGTGGGTG 52 5-8-5 1327 621566 6087 6104 TGGTGCCGGAGCTGGTGG 64 5-8-5 1328 621567 6090 6107 TGTTGGTGCCGGAGCTGG 74 5-8-5 1329 621568 6093 6110 TGCTGTTGGTGCCGGAGC 50 5-8-5 1330 621569 6096 6113 CGCTGCTGTTGGTGCCGG 47 5-8-5 1331 621570 6111 6128 GGGCGGTGGCAGCGGCGC 86 5-8-5 1332 621571 6114 6131 GGTGGGCGGTGGCAGCGG 61 5-8-5 1333 621572 6117 6134 GAAGGTGGGCGGTGGCAG 19 5-8-5 1334 621573 6120 6137 GCAGAAGGTGGGCGGTGG 22 5-8-5 1335 621574 6123 6140 GCGGCAGAAGGTGGGCGG 18 5-8-5 1336 621575 6126 6143 GCGGCGGCAGAAGGTGGG 62 5-8-5 1337 621576 6129 6146 GTGGCGGCGGCAGAAGGT 79 5-8-5 1338 621577 6132 6149 GTGGTGGCGGCGGCAGAA 70 5-8-5 1339 621578 6135 6152 GCTGTGGTGGCGGCGGCA 78 5-8-5 1340 621579 6138 6155 GTGGCTGTGGTGGCGGCG 78 5-8-5 1341 621580 6141 6158 AAGGTGGCTGTGGTGGCG 72 5-8-5 1342 613412 73879 73898 GTGATCTTCCATCACTTCGA 78 5-8-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 94 5-8-5 665 98928 98945 621504 132626 132643 AAACCCTTCTCACAAAAC 17 5-8-5 1343 621505 132812 132829 CCTCCTGGTCCCTCCTCG 12 5-8-5 1344 621506 132992 133009 CCTATCCAGACCCACTCA 10 5-8-5 1345 621507 133179 133196 CACAACCACCCCGGGACG 8 5-8-5 1346 621508 133359 133376 GGACCAGGATGGTGTTTT 44 5-8-5 1347 621509 133539 133556 GGCTTTGATAAAAATATT 8 5-8-5 1348 621510 133788 133805 CGCCTCCGGGATCAAGCA 47 5-8-5 1349 621511 133968 133985 GGTTCGCATTTAAGCTAG 28 5-8-5 1350 621512 134148 134165 ACGGTCATTCTCAGCCCT 59 5-8-5 1351 621513 134333 134350 TAACCTCCAGAGCAACTG 26 5-8-5 1352 621514 134513 134530 GGCAGGTGGGCCACAAAA 17 5-8-5 1353 621515 134696 134713 TCAAAAGGCATGCACATT 44 5-8-5 1354 621516 134876 134893 TAGTATTCCGTCGCCACG 40 5-8-5 1355 621517 135057 135074 GAGCTGTTGTTTTTATTG 7 5-8-5 1356 621518 135369 135386 GCCAGGGACTGTTTTTTG 62 5-8-5 1357

TABLE 28 Inhibition of tau mRNA by 5-8-5 MOE gapmers targeting SEQ ID NOs: 1 and 2 SEQ ID SEQ ID NO: 1 NO: 1 SEQ ID SEQ ID SEQ start stop % NO: 2 NO: 2 ID ISIS NO site site Sequence inhibition start site stop site Motif NO: 621596 n/a n/a TCACCTGATAGTCGACAG 70 292 309 5-8-5 1358 621597 n/a n/a AGTTCACCTGATAGTCGA 61 295 312 5-8-5 1359 621598 n/a n/a CAAAGTTCACCTGATAGT 44 298 315 5-8-5 1360 621599 n/a n/a GTTCAAAGTTCACCTGAT 72 301 318 5-8-5 1361 621629 n/a n/a GATTCTTTCAGGCCAGCG 42 442 459 5-8-5 1362 621630 n/a n/a GGAGATTCTTTCAGGCCA 0 445 462 5-8-5 1363 621649 n/a n/a ATCTTCCGCTGTTGGAGT 43 527 544 5-8-5 1364 621650 n/a n/a CACATCTTCCGCTGTTGG 3 530 547 5-8-5 1365 621651 n/a n/a TGTCACATCTTCCGCTGT 0 533 550 5-8-5 1366 621652 n/a n/a TGCTGTCACATCTTCCGC 30 536 553 5-8-5 1367 621653 n/a n/a AGGGTGCTGTCACATCTT 61 540 557 5-8-5 1368 621581 6144 6161 GAGAAGGTGGCTGTGGTG 14 244 261 5-8-5 1369 621582 6147 6164 GAGGAGAAGGTGGCTGTG 53 247 264 5-8-5 1370 621583 6153 6170 GCGGAGGAGGAGAAGGTG 25 253 270 5-8-5 1371 621584 6156 6173 ACAGCGGAGGAGGAGAAG 2 256 273 5-8-5 1372 621585 6159 6176 AGGACAGCGGAGGAGGAG 38 259 276 5-8-5 1373 621586 6162 6179 GAGAGGACAGCGGAGGAG 53 262 279 5-8-5 1374 621587 6165 6182 CGGGAGAGGACAGCGGAG 43 265 282 5-8-5 1375 621588 6168 6185 GGACGGGAGAGGACAGCG 69 268 285 5-8-5 1376 621589 6171 6188 CGAGGACGGGAGAGGACA 0 271 288 5-8-5 1377 621590 6174 6191 AGGCGAGGACGGGAGAGG 13 274 291 5-8-5 1378 621591 6177 6194 CAGAGGCGAGGACGGGAG 10 277 294 5-8-5 1379 621592 6180 6197 CGACAGAGGCGAGGACGG 57 280 297 5-8-5 1380 621593 6183 6200 AGTCGACAGAGGCGAGGA 61 283 300 5-8-5 1381 621594 6186 6203 GATAGTCGACAGAGGCGA 65 286 303 5-8-5 1382 621595 6189 6206 CCTGATAGTCGACAGAGG 51 289 306 5-8-5 1383 621600 73838 73855 CTGGTTCAAAGTTCACCT 52 304 321 5-8-5 1384 621601 73841 73858 ATCCTGGTTCAAAGTTCA 62 307 324 5-8-5 1385 621602 73844 73861 GCCATCCTGGTTCAAAGT 42 310 327 5-8-5 1386 621603 73847 73864 TCAGCCATCCTGGTTCAA 38 313 330 5-8-5 1387 621604 73850 73867 GGCTCAGCCATCCTGGTT 65 316 333 5-8-5 1388 621605 73867 73884 CTTCGAACTCCTGGCGGG 20 333 350 5-8-5 1389 621606 73870 73887 TCACTTCGAACTCCTGGC 51 336 353 5-8-5 1390 621607 73873 73890 CCATCACTTCGAACTCCT 68 339 356 5-8-5 1391 621608 73876 73893 CTTCCATCACTTCGAACT 28 342 359 5-8-5 1392 613412 73879 73898 GTGATCTTCCATCACTTCGA 75 345 364 5-8-5 25 621609 73879 73896 GATCTTCCATCACTTCGA 68 345 362 5-8-5 1393 621610 73882 73899 CGTGATCTTCCATCACTT 13 348 365 5-8-5 1394 621611 73906 73923 TGTCCCCCAACCCGTACG 46 372 389 5-8-5 1395 621612 73909 73926 TCCTGTCCCCCAACCCGT 54 375 392 5-8-5 1396 621613 73912 73929 CTTTCCTGTCCCCCAACC 43 378 395 5-8-5 1397 621614 73915 73932 GATCTTTCCTGTCCCCCA 71 381 398 5-8-5 1398 621615 73918 73935 CCTGATCTTTCCTGTCCC 71 384 401 5-8-5 1399 621616 73921 73938 CCCCCTGATCTTTCCTGT 54 387 404 5-8-5 1400 621617 73924 73941 AGCCCCCCTGATCTTTCC 40 390 407 5-8-5 1401 621618 73927 73944 TGTAGCCCCCCTGATCTT 35 393 410 5-8-5 1402 621619 73930 73947 TGGTGTAGCCCCCCTGAT 31 396 413 5-8-5 1403 621620 73933 73950 GCATGGTGTAGCCCCCCT 71 399 416 5-8-5 1404 621621 73936 73953 GGTGCATGGTGTAGCCCC 62 402 419 5-8-5 1405 621622 73939 73956 CTTGGTGCATGGTGTAGC 54 405 422 5-8-5 1406 621623 73942 73959 GGTCTTGGTGCATGGTGT 65 408 425 5-8-5 1407 621624 73945 73962 CTTGGTCTTGGTGCATGG 63 411 428 5-8-5 1408 621625 73948 73965 CCTCTTGGTCTTGGTGCA 49 414 431 5-8-5 1409 621626 73951 73968 CACCCTCTTGGTCTTGGT 65 417 434 5-8-5 1410 621627 73956 73973 CGTGTCACCCTCTTGGTC 49 422 439 5-8-5 1411 621628 73959 73976 GTCCGTGTCACCCTCTTG 70 425 442 5-8-5 1412 621631 83395 83412 AGATCCGTCCTCAGTGGG 48 473 490 5-8-5 1413 621632 83398 83415 CTCAGATCCGTCCTCAGT 38 476 493 5-8-5 1414 621633 83401 83418 TTCCTCAGATCCGTCCTC 11 479 496 5-8-5 1415 621634 83404 83421 CGGTTCCTCAGATCCGTC 57 482 499 5-8-5 1416 621635 83407 83424 GCCCGGTTCCTCAGATCC 38 485 502 5-8-5 1417 621636 83410 83427 AGAGCCCGGTTCCTCAGA 63 488 505 5-8-5 1418 621637 83413 83430 TTCAGAGCCCGGTTCCTC 50 491 508 5-8-5 1419 621638 83416 83433 GGTTTCAGAGCCCGGTTC 48 494 511 5-8-5 1420 621639 83419 83436 AGAGGTTTCAGAGCCCGG 41 497 514 5-8-5 1421 621640 83422 83439 ATCAGAGGTTTCAGAGCC 40 500 517 5-8-5 1422 621641 83425 83442 AGCATCAGAGGTTTCAGA 23 503 520 5-8-5 1423 621642 83428 83445 CTTAGCATCAGAGGTTTC 50 506 523 5-8-5 1424 621643 83431 83448 GCTCTTAGCATCAGAGGT 69 509 526 5-8-5 1425 621644 83434 83451 AGTGCTCTTAGCATCAGA 80 512 529 5-8-5 1426 621645 83437 83454 TGGAGTGCTCTTAGCATC 50 515 532 5-8-5 1427 621646 83440 83457 TGTTGGAGTGCTCTTAGC 59 518 535 5-8-5 1428 621647 83443 83460 CGCTGTTGGAGTGCTCTT 70 521 538 5-8-5 1429 621648 83446 83463 TTCCGCTGTTGGAGTGCT 52 524 541 5-8-5 1430 621654 85904 85921 CTAAGGGTGCTGTCACAT 64 543 560 5-8-5 1431 621655 85907 85924 CCACTAAGGGTGCTGTCA 44 546 563 5-8-5 1432 621656 85910 85927 CATCCACTAAGGGTGCTG 55 549 566 5-8-5 1433 621657 85913 85930 CCTCATCCACTAAGGGTG 41 552 569 5-8-5 1434 620887 98891 98908 GTTTTCAAACACACCTTC 81 n/a n/a 5-8-5 665 98928 98945 n/a n/a

TABLE 29 Inhibition of Tau mRNA by 5-8-5 MOE gapmers targeting SEQ ID NOs: 1 and 2 SEQ ID SEQ ID SEQ ID SEQ ID SEQ NO: 1 NO: 1 % NO: 2 NO: 2 ID ISIS NO start site stop site Sequence inhibition start site stop site Motif NO 621672 n/a n/a GCTGTGGTTCCTTCTGGG 51 613 630 5-8-5 1435 621676 n/a n/a TCAGGCTCTTGGGTCACG 0 685 702 5-8-5 1436 621677 n/a n/a CTTTCAGGCTCTTGGGTC 0 688 705 5-8-5 1437 621678 n/a n/a CCACTTTCAGGCTCTTGG 51 691 708 5-8-5 1438 613412 73879 73898 GTGATCTTCCATCACTTCGA 79 345 364 5-10-5 25 621658 85916 85933 CTCCCTCATCCACTAAGG 27 555 572 5-8-5 1439 621659 85919 85936 GAGCTCCCTCATCCACTA 35 558 575 5-8-5 1440 621660 85922 85939 CGGGAGCTCCCTCATCCA 25 561 578 5-8-5 1441 621661 85925 85942 TGCCGGGAGCTCCCTCAT 40 564 581 5-8-5 1442 621662 85928 85945 GCTTGCCGGGAGCTCCCT 47 567 584 5-8-5 1443 621663 85931 85948 CCTGCTTGCCGGGAGCTC 46 570 587 5-8-5 1444 621664 85934 85951 CAGCCTGCTTGCCGGGAG 45 573 590 5-8-5 1445 621665 85937 85954 CGGCAGCCTGCTTGCCGG 1 576 593 5-8-5 1446 621666 85940 85957 GCGCGGCAGCCTGCTTGC 31 579 596 5-8-5 1447 621667 85943 85960 GCTGCGCGGCAGCCTGCT 50 582 599 5-8-5 1448 621668 85962 85979 TCTGGGATCTCCGTGTGG 39 601 618 5-8-5 1449 621669 85965 85982 CCTTCTGGGATCTCCGTG 56 604 621 5-8-5 1450 621670 85968 85985 GTTCCTTCTGGGATCTCC 79 607 624 5-8-5 1451 621671 85971 85988 GTGGTTCCTTCTGGGATC 49 610 627 5-8-5 1452 621673 89894 89911 CAATGCCTGCTTCTTCAG 31 630 647 5-8-5 1453 621674 89899 89916 GTCTCCAATGCCTGCTTC 43 635 652 5-8-5 1454 621675 89902 89919 GGTGTCTCCAATGCCTGC 74 638 655 5-8-5 1455 621679 94695 94712 TTACCACTTTCAGGCTCT 63 694 711 5-8-5 1456 621680 94700 94717 CCACCTTACCACTTTCAG 28 699 716 5-8-5 1457 621681 94703 94720 GGACCACCTTACCACTTT 62 702 719 5-8-5 1458 621682 94706 94723 CCTGGACCACCTTACCAC 33 705 722 5-8-5 1459 621683 94709 94726 CTTCCTGGACCACCTTAC 18 708 725 5-8-5 1460 621684 94712 94729 AGCCTTCCTGGACCACCT 61 711 728 5-8-5 1461 621685 94715 94732 GGAAGCCTTCCTGGACCA 54 714 731 5-8-5 1462 621686 94718 94735 GGAGGAAGCCTTCCTGGA 24 717 734 5-8-5 1463 621687 94721 94738 CTCGGAGGAAGCCTTCCT 48 720 737 5-8-5 1464 621688 94724 94741 GCTCTCGGAGGAAGCCTT 27 723 740 5-8-5 1465 621689 94727 94744 CTGGCTCTCGGAGGAAGC 19 726 743 5-8-5 1466 621690 94730 94747 GGCCTGGCTCTCGGAGGA 46 729 746 5-8-5 1467 621691 94749 94766 TGGTGGCTCAGACCTGGG 49 748 765 5-8-5 1468 621692 94752 94769 AGCTGGTGGCTCAGACCT 50 751 768 5-8-5 1469 621693 94755 94772 ATGAGCTGGTGGCTCAGA 17 754 771 5-8-5 1470 621694 94758 94775 GACATGAGCTGGTGGCTC 54 757 774 5-8-5 1471 621695 94761 94778 CCGGACATGAGCTGGTGG 26 760 777 5-8-5 1472 621696 94764 94781 ATGCCGGACATGAGCTGG 12 763 780 5-8-5 1473 621697 94767 94784 GGCATGCCGGACATGAGC 11 766 783 5-8-5 1474 621698 94770 94787 CCAGGCATGCCGGACATG 41 769 786 5-8-5 1475 621699 94773 94790 GCCCCAGGCATGCCGGAC 57 772 789 5-8-5 1476 621700 94776 94793 GGAGCCCCAGGCATGCCG 47 775 792 5-8-5 1477 621701 94793 94810 GGCCCTCAGGCAGGAGGG 0 792 809 5-8-5 1478 621702 94825 94842 TGTCCCCGAAGGTTGGCG 50 824 841 5-8-5 1479 621703 94828 94845 TCCTGTCCCCGAAGGTTG 38 827 844 5-8-5 1480 621704 94831 94848 AGGTCCTGTCCCCGAAGG 41 830 847 5-8-5 1481 621705 94834 94851 CTCAGGTCCTGTCCCCGA 60 833 850 5-8-5 1482 621706 94837 94854 GTCCTCAGGTCCTGTCCC 0 836 853 5-8-5 1483 621707 94840 94857 TGTGTCCTCAGGTCCTGT 23 839 856 5-8-5 1484 621708 94843 94860 CTCTGTGTCCTCAGGTCC 55 842 859 5-8-5 1485 621709 94846 94863 GCCCTCTGTGTCCTCAGG 35 845 862 5-8-5 1486 621710 94849 94866 GCCGCCCTCTGTGTCCTC 45 848 865 5-8-5 1487 621711 94852 94869 GCGGCCGCCCTCTGTGTC 0 851 868 5-8-5 1488 621712 94855 94872 GTGGCGGCCGCCCTCTGT 0 854 871 5-8-5 1489 621713 94875 94892 TGCTTGAGCAGCTCAGGG 67 874 891 5-8-5 1490 621714 94878 94895 TGGTGCTTGAGCAGCTCA 51 877 894 5-8-5 1491 621715 94882 94899 AAGCTGGTGCTTGAGCAG 27 881 898 5-8-5 1492 621716 94885 94902 TAGAAGCTGGTGCTTGAG 14 884 901 5-8-5 1493 621717 94888 94905 TCCTAGAAGCTGGTGCTT 0 887 904 5-8-5 1494 621718 94891 94908 GTCTCCTAGAAGCTGGTG 45 890 907 5-8-5 1495 621719 94894 94911 CAGGTCTCCTAGAAGCTG 47 893 910 5-8-5 1496 621720 94897 94914 GTGCAGGTCTCCTAGAAG 41 896 913 5-8-5 1497 621721 94900 94917 CTGGTGCAGGTCTCCTAG 36 899 916 5-8-5 1498 621722 94903 94920 CTCCTGGTGCAGGTCTCC 41 902 919 5-8-5 1499 621723 94906 94923 CCCCTCCTGGTGCAGGTC 39 905 922 5-8-5 1500 621724 94909 94926 CGGCCCCTCCTGGTGCAG 19 908 925 5-8-5 1501 621725 94912 94929 CGGCGGCCCCTCCTGGTG 22 911 928 5-8-5 1502 621726 94915 94932 CAGCGGCGGCCCCTCCTG 35 914 931 5-8-5 1503 621727 94918 94935 CTTCAGCGGCGGCCCCTC 46 917 934 5-8-5 1504 621728 94921 94938 CCCCTTCAGCGGCGGCCC 36 920 937 5-8-5 1505 621729 94924 94941 TGCCCCCTTCAGCGGCGG 17 923 940 5-8-5 1506 621730 94927 94944 CCCTGCCCCCTTCAGCGG 15 926 943 5-8-5 1507 621731 94930 94947 GCCCCCTGCCCCCTTCAG 27 929 946 5-8-5 1508 621732 94933 94950 TTTGCCCCCTGCCCCCTT 49 932 949 5-8-5 1509 621733 94936 94953 CTCTTTGCCCCCTGCCCC 22 935 952 5-8-5 1510 621734 94939 94956 CCTCTCTTTGCCCCCTGC 28 938 955 5-8-5 1511 620887 98891 98908 GTTTTCAAACACACCTTC 90 n/a n/a 5-8-5 665 98928 98945 n/a n/a

TABLE 30 Inhibition of tau mRNA by 5-10-5 MOE and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 SEQ ID SEQ ID NO: 1 start NO: 1 stop % SEQ ISIS NO site site Sequence inhibition Motif ID NO: 613412 73879 73898 GTGATCTTCCATCACTTCGA 82 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 92 5-8-5 665 98928 98945 621735 94942 94959 CGGCCTCTCTTTGCCCCC 34 5-8-5 1512 621736 94945 94962 CCCCGGCCTCTCTTTGCC 27 5-8-5 1513 621737 94948 94965 GCTCCCCGGCCTCTCTTT 44 5-8-5 1514 621738 94951 94968 CTTGCTCCCCGGCCTCTC 38 5-8-5 1515 621739 94954 94971 CTCCTTGCTCCCCGGCCT 37 5-8-5 1516 621740 94957 94974 CTCCTCCTTGCTCCCCGG 33 5-8-5 1517 621741 94960 94977 CACCTCCTCCTTGCTCCC 37 5-8-5 1518 621742 94963 94980 ATCCACCTCCTCCTTGCT 32 5-8-5 1519 621743 94967 94984 CTTCATCCACCTCCTCCT 51 5-8-5 1520 621744 94970 94987 GGTCTTCATCCACCTCCT 61 5-8-5 1521 621745 94973 94990 CGCGGTCTTCATCCACCT 71 5-8-5 1522 621746 94976 94993 CGTCGCGGTCTTCATCCA 48 5-8-5 1523 621747 95038 95055 CCGCCCATCTTGGGCTGG 12 5-8-5 1524 621748 95041 95058 AGGCCGCCCATCTTGGGC 18 5-8-5 1525 621749 95044 95061 GGGAGGCCGCCCATCTTG 21 5-8-5 1526 621750 95060 95077 CTCTGGCGGCTGTCTGGG 47 5-8-5 1527 621751 95063 95080 CTTCTCTGGCGGCTGTCT 41 5-8-5 1528 621752 95066 95083 TGGCTTCTCTGGCGGCTG 53 5-8-5 1529 621753 95069 95086 TGGTGGCTTCTCTGGCGG 26 5-8-5 1530 621754 95072 95089 TGCTGGTGGCTTCTCTGG 57 5-8-5 1531 621755 95075 95092 GGATGCTGGTGGCTTCTC 65 5-8-5 1532 621756 95078 95095 CTGGGATGCTGGTGGCTT 66 5-8-5 1533 621757 95081 95098 AGCCTGGGATGCTGGTGG 51 5-8-5 1534 621758 95084 95101 GGAAGCCTGGGATGCTGG 57 5-8-5 1535 621759 95092 95109 CTCCGCTGGGAAGCCTGG 48 5-8-5 1536 621760 95095 95112 ACCCTCCGCTGGGAAGCC 38 5-8-5 1537 621761 95098 95115 GGCACCCTCCGCTGGGAA 54 5-8-5 1538 621762 95101 95118 GATGGCACCCTCCGCTGG 27 5-8-5 1539 621763 95121 95138 AGGAAATCCACAGGGAGG 15 5-8-5 1540 621764 95124 95141 GAGAGGAAATCCACAGGG 29 5-8-5 1541 621765 95127 95144 TTGGAGAGGAAATCCACA 45 5-8-5 1542 621766 95130 95147 ACTTTGGAGAGGAAATCC 38 5-8-5 1543 621767 95134 95151 GGAAACTTTGGAGAGGAA 56 5-8-5 1544 621768 95137 95154 TGTGGAAACTTTGGAGAG 26 5-8-5 1545 621769 95140 95157 CTCTGTGGAAACTTTGGA 69 5-8-5 1546 621770 95143 95160 GATCTCTGTGGAAACTTT 62 5-8-5 1547 621771 95146 95163 TGGGATCTCTGTGGAAAC 61 5-8-5 1548 621772 95149 95166 GGCTGGGATCTCTGTGGA 47 5-8-5 1549 621773 95152 95169 TGAGGCTGGGATCTCTGT 32 5-8-5 1550 621774 95155 95172 CTCTGAGGCTGGGATCTC 64 5-8-5 1551 621775 95161 95178 GTCGGGCTCTGAGGCTGG 42 5-8-5 1552 621776 95164 95181 CCCGTCGGGCTCTGAGGC 34 5-8-5 1553 621777 95174 95191 CTACACTGGGCCCGTCGG 31 5-8-5 1554 621778 95177 95194 GCCCTACACTGGGCCCGT 48 5-8-5 1555 621779 95180 95197 CCCGCCCTACACTGGGCC 35 5-8-5 1556 621780 95183 95200 TGGCCCGCCCTACACTGG 51 5-8-5 1557 621781 95186 95203 CTTTGGCCCGCCCTACAC 4 5-8-5 1558 621782 95189 95206 GCCCTTTGGCCCGCCCTA 52 5-8-5 1559 621783 95192 95209 CCTGCCCTTTGGCCCGCC 42 5-8-5 1560 621784 95195 95212 CATCCTGCCCTTTGGCCC 49 5-8-5 1561 621785 95198 95215 GGGCATCCTGCCCTTTGG 15 5-8-5 1562 621786 95235 95252 TTGGGTGTGATTTCCACG 52 5-8-5 1563 621787 95253 95270 GCCTGCTCCTTCTGCACG 44 5-8-5 1564 621788 95256 95273 TGCGCCTGCTCCTTCTGC 64 5-8-5 1565 621789 95259 95276 GAGTGCGCCTGCTCCTTC 70 5-8-5 1566 621790 95262 95279 TCCGAGTGCGCCTGCTCC 53 5-8-5 1567 621791 95265 95282 TCCTCCGAGTGCGCCTGC 54 5-8-5 1568 621792 95268 95285 TGCTCCTCCGAGTGCGCC 52 5-8-5 1569 621793 95271 95288 AAATGCTCCTCCGAGTGC 72 5-8-5 1570 621794 95274 95291 CCCAAATGCTCCTCCGAG 75 5-8-5 1571 621795 95277 95294 CTTCCCAAATGCTCCTCC 47 5-8-5 1572 621796 95280 95297 GCCCTTCCCAAATGCTCC 58 5-8-5 1573 621797 95283 95300 GCAGCCCTTCCCAAATGC 39 5-8-5 1574 621798 95286 95303 AATGCAGCCCTTCCCAAA 61 5-8-5 1575 621799 95291 95308 CTGGAAATGCAGCCCTTC 64 5-8-5 1576 621800 95294 95311 CCCCTGGAAATGCAGCCC 49 5-8-5 1577 621801 95297 95314 GGGCCCCTGGAAATGCAG 29 5-8-5 1578 621802 95313 95330 TCTGGCCCCTCTCCAGGG 41 5-8-5 1579 621803 95316 95333 GCCTCTGGCCCCTCTCCA 42 5-8-5 1580 621804 95319 95336 CGGGCCTCTGGCCCCTCT 11 5-8-5 1581 621805 95322 95339 CCCCGGGCCTCTGGCCCC 13 5-8-5 1582 621806 95341 95358 GTCCTCTCCCAAAGAGGG 25 5-8-5 1583 621807 95344 95361 TGTGTCCTCTCCCAAAGA 20 5-8-5 1584 621808 95347 95364 TTTTGTGTCCTCTCCCAA 66 5-8-5 1585 621809 95365 95382 CTCTGGAAGGTCAGCCTC 53 5-8-5 1586 621810 95368 95385 GGGCTCTGGAAGGTCAGC 72 5-8-5 1587 621811 95371 95388 AGAGGGCTCTGGAAGGTC 57 5-8-5 1588

TABLE 31 Inhibition of tau mRNA by 5-10-5 and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 SEQ ID SEQ ID ISIS NO: 1 start NO: 1 stop % SEQ ID NO site site Sequence inhibition Motif NO: 613412 73879 73898 GTGATCTTCCATCACTTCGA 88 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 96 5-8-5 665 98928 98945 623737 111219 111236 CCTCCACCCAGCATGGTG 56 5-8-5 1589 623738 111267 111284 TCAAGTCACCCTTTCTCC 78 5-8-5 1590 623739 111368 111385 TTCAGACAATTTTTCTAG 66 5-8-5 1591 623740 111629 111646 TCAGACTCCGCCAGCTTT 79 5-8-5 1592 623741 111678 111695 TCTTTTCCCTTTGGTGTT 34 5-8-5 1593 623742 111753 111770 TTTCCCCCAATGATTTGC 77 5-8-5 1594 623743 112016 112033 CCACGACTCCCACAAGAT 32 5-8-5 1595 623744 112213 112230 TGAGGTTGAGGGTAGGTG 50 5-8-5 1596 623745 112219 112236 AGTGGTTGAGGTTGAGGG 31 5-8-5 1597 623746 112304 112321 CAGGCACTTGGAAACTGC 90 5-8-5 1598 623747 112877 112894 TTGTACTCTTTTTCCCCT 65 5-8-5 1599 623748 112949 112966 TTAGGAGTGCAAGGTTGT 49 5-8-5 1600 623749 113352 113369 TCCTACATTCCTCCTGCT 67 5-8-5 1601 623750 113523 113540 GTTGCAGTGTTCCACTAT 90 5-8-5 1602 623751 113783 113800 CCAGCAGATGCCGACAGC 74 5-8-5 1603 623752 113809 113826 GGGCCCTCACCCCTGCTT 14 5-8-5 1604 623753 113830 113847 CCAAGAAGGGCTGCTGAG 8 5-8-5 1605 623754 114267 114284 AAGCAGCCACCCAGGCTG 17 5-8-5 1606 623755 114739 114756 TATGAGGAGGGAGGAAAG 13 5-8-5 1607 623756 115242 115259 CTGACATCTCAGCCCAAG 93 5-8-5 1608 621425 115335 115352 GGTACAGGAAAAGATGCC 77 5-8-5 1202 623757 115522 115539 CAAGGTGATCAGCTTAGG 82 5-8-5 1609 623758 115526 115543 AGTCCAAGGTGATCAGCT 89 5-8-5 1610 623759 115547 115564 CCCACACAAGCCTCCTCT 66 5-8-5 1611 623760 115791 115808 TTTGTTTGGGTTCAGTTC 59 5-8-5 1612 623761 115803 115820 CCTTGATCTGGTTTTGTT 77 5-8-5 1613 623762 115993 116010 ATCAAGGGAGAGAACGAT 52 5-8-5 1614 623763 116229 116246 AAAAGGCAACAAGCCTTT 0 5-8-5 1615 623764 116769 116786 CGAGGCAAAAAGGGAAAT 45 5-8-5 1616 623765 116791 116808 TAACTTTGAGCTTCCTGG 65 5-8-5 1617 623766 116944 116961 TAAGGCTTAGAAGATTTG 45 5-8-5 1618 623767 116978 116995 GACGATCATTTTTTATTC 72 5-8-5 1619 623768 116983 117000 GGCTTGACGATCATTTTT 77 5-8-5 1620 623769 117421 117438 AGATCTCAGAGTTTGTAG 68 5-8-5 1621 623770 117472 117489 TCGCTGAGGTTCCGCAAC 37 5-8-5 1622 623771 117489 117506 AGGAGGCCATGTGAGGCT 61 5-8-5 1623 623772 117547 117564 TGGATAAATCTCTAGTGC 54 5-8-5 1624 623773 117564 117581 CACTCCTCCTTTAGAGAT 38 5-8-5 1625 623774 117599 117616 TTGTAATTCCTTCTCTGG 65 5-8-5 1626 623775 117971 117988 TAACCATCCCTACTTTCT 42 5-8-5 1627 623776 119098 119115 ACTGAAATCGCTCCTTTA 58 5-8-5 1628 623777 119107 119124 CCAATTATGACTGAAATC 32 5-8-5 1629 623778 119313 119330 GGTCAAAGTTATAAATAT 57 5-8-5 1630 623779 119431 119448 GGATTTGATGCAAAGAAA 70 5-8-5 1631 623780 119740 119757 GCGCAACCCAGCTACTCG 7 5-8-5 1632 621445 119880 119897 TACAGTGGCTCCTGTAAT 28 5-8-5 1222 623781 119964 119981 CTGTATACCTATACTTGG 34 5-8-5 1633 623782 120043 120060 CCGTTTTCTTACCACCCT 92 5-8-5 1634 623783 120177 120194 TTACTTGTCTGAATCTTC 83 5-8-5 1635 623784 120204 120221 TTCATCCCGTTTTTTTTC 54 5-8-5 1636 623785 120309 120326 GACCTCAGTGGCTCTTTT 68 5-8-5 1637 623786 120513 120530 TGTCCAGGATAGTTTGAA 44 5-8-5 1638 623787 121033 121050 GCTGAGCAAATGCTCCCG 54 5-8-5 1639 623788 121194 121211 CTGACCCCGTAGGCAGGA 29 5-8-5 1640 623789 121478 121495 ACCAGCCTGAGGTCAAGT 73 5-8-5 1641 623790 121538 121555 CAGGTCCTCACTTCACAA 42 5-8-5 1642 623791 121767 121784 CCGCCTGCTTGCTCGCAA 92 5-8-5 1643 623792 121787 121804 AGTGACACGCCACCCTGG 80 5-8-5 1644 623793 121797 121814 AAAAAGGATGAGTGACAC 28 5-8-5 1645 623794 121807 121824 GGTAGCCAGAAAAAAGGA 49 5-8-5 1646 623795 121812 121829 CCTTTGGTAGCCAGAAAA 63 5-8-5 1647 623796 121817 121834 CTGCACCTTTGGTAGCCA 87 5-8-5 1648 623797 121822 121839 ATTATCTGCACCTTTGGT 67 5-8-5 1649 623798 121824 121841 TAATTATCTGCACCTTTG 49 5-8-5 1650 623799 121905 121922 CACTGCCGCCTCCCGGGA 65 5-8-5 1651 623800 121909 121926 CTCACACTGCCGCCTCCC 75 5-8-5 1652 623801 121911 121928 TACTCACACTGCCGCCTC 54 5-8-5 1653 623679 121913 121930 GGTACTCACACTGCCGCC 81 5-8-5 1654 623802 121915 121932 AAGGTACTCACACTGCCG 69 5-8-5 1655 623803 121920 121937 GTGTGAAGGTACTCACAC 18 5-8-5 1656 623804 121942 121959 GCCACAGCACGGCGCATG 80 5-8-5 1657 623805 121952 121969 AATAATTCAAGCCACAGC 88 5-8-5 1658 623806 121962 121979 ACCACTTCCTAATAATTC 39 5-8-5 1659 623807 121972 121989 ACGCACTCACACCACTTC 83 5-8-5 1660 623808 121982 121999 TCGCAAGTGTACGCACTC 94 5-8-5 1661 623809 121992 122009 ATGCAGTGTCTCGCAAGT 93 5-8-5 1662 623810 122227 122244 TGATGGGTATTCTCAGCT 77 5-8-5 1663

TABLE 32 Inhibition of tau mRNA by 5-10-5 MOE and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 SEQ ID SEQ ID SEQ ISIS NO: 1 start NO: 1 stop % ID NO site site Sequence inhibition Motif NO: 613412 73879 73898 GTGATCTTCCATCACTTCGA 87 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 83 5-8-5 665 98928 98945 622186 5897 5914 CCGCTCGGCCGTCCGGCG 6 5-8-5 1664 622187 5900 5917 CTGCCGCTCGGCCGTCCG 6 5-8-5 1665 622174 103089 103106 CCAGAGCTGGGTGGTGTC 12 5-8-5 1666 622182 125760 125777 TGTAGACTATTTGCACCT 74 5-8-5 1667 622120 135855 135872 ATCACTGATTTTGAAGTC 36 5-8-5 1668 622121 135858 135875 CCCATCACTGATTTTGAA 59 5-8-5 1669 622122 135861 135878 ACTCCCATCACTGATTTT 70 5-8-5 1670 622123 135864 135881 CTTACTCCCATCACTGAT 41 5-8-5 1671 622124 135867 135884 GCTCTTACTCCCATCACT 82 5-8-5 1672 622125 135870 135887 TTTGCTCTTACTCCCATC 87 5-8-5 1673 622126 135873 135890 AAATTTGCTCTTACTCCC 62 5-8-5 1674 622127 135876 135893 ATGAAATTTGCTCTTACT 70 5-8-5 1675 622128 135879 135896 AAGATGAAATTTGCTCTT 55 5-8-5 1676 622129 135882 135899 GGAAAGATGAAATTTGCT 84 5-8-5 1677 622130 135885 135902 TTTGGAAAGATGAAATTT 12 5-8-5 1678 622131 135891 135908 CATCAATTTGGAAAGATG 43 5-8-5 1679 622132 135894 135911 ACCCATCAATTTGGAAAG 18 5-8-5 1680 622133 135897 135914 CCCACCCATCAATTTGGA 34 5-8-5 1681 622134 135900 135917 TAGCCCACCCATCAATTT 17 5-8-5 1682 622135 135903 135920 TACTAGCCCACCCATCAA 14 5-8-5 1683 622136 135906 135923 TATTACTAGCCCACCCAT 24 5-8-5 1684 622137 135909 135926 TTTTATTACTAGCCCACC 24 5-8-5 1685 622138 135912 135929 ATATTTTATTACTAGCCC 33 5-8-5 1686 622139 135915 135932 TAAATATTTTATTACTAG 0 5-8-5 1687 622140 135918 135935 TTTTAAATATTTTATTAC 0 5-8-5 1688 622141 135926 135943 ATGTTTTTTTTTAAATAT 0 5-8-5 1689 622142 135939 135956 GCCATGTTTTTGAATGTT 60 5-8-5 1690 622143 135942 135959 GTGGCCATGTTTTTGAAT 28 5-8-5 1691 622144 135945 135962 GATGTGGCCATGTTTTTG 63 5-8-5 1692 622145 135948 135965 TTGGATGTGGCCATGTTT 78 5-8-5 1693 622146 135951 135968 ATGTTGGATGTGGCCATG 71 5-8-5 1694 622147 135954 135971 GAAATGTTGGATGTGGCC 72 5-8-5 1695 622148 135957 135974 GAGGAAATGTTGGATGTG 55 5-8-5 1696 622149 135960 135977 CCTGAGGAAATGTTGGAT 77 5-8-5 1697 622150 135963 135980 TTGCCTGAGGAAATGTTG 55 5-8-5 1698 622151 139811 139828 CCAAATTCACTTTTACAG 54 5-8-5 1699 622152 139813 139830 TTCCAAATTCACTTTTAC 67 5-8-5 1700 622153 139815 139832 ATTTCCAAATTCACTTTT 52 5-8-5 1701 622154 139817 139834 TTATTTCCAAATTCACTT 52 5-8-5 1702 622155 139819 139836 CTTTATTTCCAAATTCAC 61 5-8-5 1703 622156 139821 139838 AACTTTATTTCCAAATTC 26 5-8-5 1704 622157 139823 139840 ATAACTTTATTTCCAAAT 23 5-8-5 1705 622158 139826 139843 GTAATAACTTTATTTCCA 65 5-8-5 1706 622159 139828 139845 GAGTAATAACTTTATTTC 25 5-8-5 1707 622160 139830 139847 CAGAGTAATAACTTTATT 9 5-8-5 1708 622161 139832 139849 ATCAGAGTAATAACTTTA 23 5-8-5 1709 622162 139834 139851 TAATCAGAGTAATAACTT 14 5-8-5 1710 622163 139836 139853 TTTAATCAGAGTAATAAC 4 5-8-5 1711

TABLE 33 Inhibition of Tau mRNA by 5-8-5 MOE gapmers targeting SEQ ID NO: 5 and 6 SEQ SEQ SEQ SEQ ID ID ID ID NO: 5 NO: 5 NO: 6 NO: 6 SEQ ISIS Start Stop % Start Stop ID NO Site Site Sequence inhibition Site Site Motif NO 622164 n/a n/a GCTTCCGCTGTTGGAGTG 22 526 543 5-8-5 1712 622165 n/a n/a TCAGCTTCCGCTGTTGGA 15 529 546 5-8-5 1713 622166 n/a n/a TCTTCAGCTTCCGCTGTT 0 532 549 5-8-5 1714 622167 n/a n/a GCTTCTTCAGCTTCCGCT 43 535 552 5-8-5 1715 622171 n/a n/a GCCCCCTTGGCTTTTTTG 16 652 669 5-8-5 1716 622172 n/a n/a TCAGCCCCCTTGGCTTTT 15 655 672 5-8-5 1717 622173 n/a n/a ACCATCAGCCCCCTTGGC 49 659 676 5-8-5 1718 622175 n/a n/a TCACCAGAGCTGGGTGGT 15 778 795 5-8-5 1719 622176 n/a n/a GGTTCACCAGAGCTGGGT 80 781 798 5-8-5 1720 622177 n/a n/a GGAGGTTCACCAGAGCTG 79 784 801 5-8-5 1721 622178 n/a n/a TTTGGAGGTTCACCAGAG 45 787 804 5-8-5 1722 622179 n/a n/a GATTTTGGAGGTTCACCA 68 790 807 5-8-5 1723 622180 n/a n/a TTGCACCTTCCCGCCTCC 19 1046 1063 5-8-5 1724 622181 n/a n/a AGACTATTTGCACCTTCC 70 1053 1070 5-8-5 1725 622183 3 20 CCCTTCGCGGTCCCTTCG 0 n/a n/a 5-8-5 1726 622184 6 23 CTGCCCTTCGCGGTCCCT 12 n/a n/a 5-8-5 1727 622185 9 26 GCGCTGCCCTTCGCGGTC 0 n/a n/a 5-8-5 1728 622188 424 441 GTCCCCCAAACCCGTACG 15 n/a n/a 5-8-5 1729 622189 427 444 CCTGTCCCCCAAACCCGT 45 n/a n/a 5-8-5 1730 622190 430 447 TTTCCTGTCCCCCAAACC 57 n/a n/a 5-8-5 1731 622191 436 453 TTGATCTTTCCTGTCCCC 74 n/a n/a 5-8-5 1732 622192 439 456 CCCTTGATCTTTCCTGTC 69 n/a n/a 5-8-5 1733 622193 442 459 GCCCCCTTGATCTTTCCT 7 n/a n/a 5-8-5 1734 622194 445 462 GTAGCCCCCTTGATCTTT 3 n/a n/a 5-8-5 1735 622195 448 465 GGTGTAGCCCCCTTGATC 8 n/a n/a 5-8-5 1736 622196 451 468 CATGGTGTAGCCCCCTTG 83 n/a n/a 5-8-5 1737 622168 564 581 ATGCGAGCTTGGGTCACG 65 598 615 5-8-5 1738 622169 567 584 ACCATGCGAGCTTGGGTC 62 601 618 5-8-5 1739 622170 570 587 CTGACCATGCGAGCTTGG 91 604 621 5-8-5 1740

TABLE 34 Inhibition of tau mRNA by 5-10-5 MOE and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 and 2 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ISIS NO: 1 NO: 1 % NO: 2 NO: 2 ID NO start site stop site Sequence inhibition start site stop site Motif NO: 613412 73879 73898 GTGATCTTCCATCACTTCGA 91 345 364 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 97 n/a n/a 5-8-5 665 98928 98945 n/a n/a 621832 n/a n/a GAGCTTTGAGTTGAGGGA 14 1434 1451 5-8-5 1741 621833 n/a n/a TGCGAGCTTTGAGTTGAG 0 1437 1454 5-8-5 1742 621834 n/a n/a CCATGCGAGCTTTGAGTT 10 1440 1457 5-8-5 1743 621835 n/a n/a TGACCATGCGAGCTTTGA 83 1443 1460 5-8-5 1744 621836 n/a n/a TACTGACCATGCGAGCTT 91 1446 1463 5-8-5 1745 621848 n/a n/a GATGTCTTGGCTTTTTTG 3 1492 1509 5-8-5 1746 621849 n/a n/a GTGTGGATGTCTTGGCTT 15 1497 1514 5-8-5 1747 621812 95374 95391 TTCAGAGGGCTCTGGAAG 0 1373 1390 5-8-5 1748 621813 95377 95394 CTTTTCAGAGGGCTCTGG 44 1376 1393 5-8-5 1749 621814 95380 95397 CTGCTTTTCAGAGGGCTC 56 1379 1396 5-8-5 1750 621815 95383 95400 AGGCTGCTTTTCAGAGGG 38 1382 1399 5-8-5 1751 621816 95386 95403 AGCAGGCTGCTTTTCAGA 40 1385 1402 5-8-5 1752 621817 95389 95406 AGCAGCAGGCTGCTTTTC 26 1388 1405 5-8-5 1753 621818 95392 95409 AGCAGCAGCAGGCTGCTT 12 1391 1408 5-8-5 1754 621819 95395 95412 CGGAGCAGCAGCAGGCTG 23 1394 1411 5-8-5 1755 621820 95398 95415 CCGCGGAGCAGCAGCAGG 34 1397 1414 5-8-5 1756 621821 95401 95418 CCCCCGCGGAGCAGCAGC 50 1400 1417 5-8-5 1757 621822 95404 95421 CTTCCCCCGCGGAGCAGC 48 1403 1420 5-8-5 1758 621823 95407 95424 GGGCTTCCCCCGCGGAGC 0 1406 1423 5-8-5 1759 621824 95410 95427 GACGGGCTTCCCCCGCGG 7 1409 1426 5-8-5 1760 621825 95413 95430 GCTGACGGGCTTCCCCCG 36 1412 1429 5-8-5 1761 621826 95416 95433 CCGGCTGACGGGCTTCCC 54 1415 1432 5-8-5 1762 621827 95419 95436 GACCCGGCTGACGGGCTT 7 1418 1435 5-8-5 1763 621828 95423 95440 GAGGGACCCGGCTGACGG 6 1422 1439 5-8-5 1764 621829 95426 95443 GTTGAGGGACCCGGCTGA 27 1425 1442 5-8-5 1765 621830 95429 95446 TGAGTTGAGGGACCCGGC 50 1428 1445 5-8-5 1766 621831 95432 95449 CTTTGAGTTGAGGGACCC 50 1431 1448 5-8-5 1767 621837 98559 98576 TTTTACTGACCATGCGAG 77 1449 1466 5-8-5 1768 621838 98562 98579 TGCTTTTACTGACCATGC 95 1452 1469 5-8-5 1769 621839 98565 98582 CTTTGCTTTTACTGACCA 85 1455 1472 5-8-5 1770 621840 98568 98585 CGTCTTTGCTTTTACTGA 66 1458 1475 5-8-5 1771 621841 98571 98588 TCCCGTCTTTGCTTTTAC 79 1461 1478 5-8-5 1772 621842 98575 98592 CCAGTCCCGTCTTTGCTT 87 1465 1482 5-8-5 1773 621843 98578 98595 CTTCCAGTCCCGTCTTTG 64 1468 1485 5-8-5 1774 621844 98581 98598 TCGCTTCCAGTCCCGTCT 88 1471 1488 5-8-5 1775 621845 98584 98601 TCATCGCTTCCAGTCCCG 90 1474 1491 5-8-5 1776 621846 98587 98604 TTGTCATCGCTTCCAGTC 88 1477 1494 5-8-5 1777 621847 98590 98607 TTTTTGTCATCGCTTCCA 80 1480 1497 5-8-5 1778 621850 101406 101423 GGTTTTAGCAGAGGAACG 75 1514 1531 5-8-5 1779 621851 101409 101426 CAAGGTTTTAGCAGAGGA 83 1517 1534 5-8-5 1780 621852 101412 101429 TTTCAAGGTTTTAGCAGA 35 1520 1537 5-8-5 1781 621853 101415 101432 ATTTTTCAAGGTTTTAGC 79 1523 1540 5-8-5 1782 621854 101418 101435 CCTATTTTTCAAGGTTTT 71 1526 1543 5-8-5 1783 621855 101425 101442 GGCAAGGCCTATTTTTCA 83 1533 1550 5-8-5 1784 621856 101428 101445 TAAGGCAAGGCCTATTTT 23 1536 1553 5-8-5 1785 621857 101431 101448 GGCTAAGGCAAGGCCTAT 69 1539 1556 5-8-5 1786 621858 101457 101474 TGAGCTACCAGGAGTGGG 75 1565 1582 5-8-5 1787 621859 101460 101477 GTCTGAGCTACCAGGAGT 78 1568 1585 5-8-5 1788 621860 101463 101480 AGGGTCTGAGCTACCAGG 82 1571 1588 5-8-5 1789 621861 101466 101483 CAGAGGGTCTGAGCTACC 88 1574 1591 5-8-5 1790 621862 101469 101486 GATCAGAGGGTCTGAGCT 72 1577 1594 5-8-5 1791 621863 101472 101489 TTGGATCAGAGGGTCTGA 83 1580 1597 5-8-5 1792 621864 101478 101495 GGAGGGTTGGATCAGAGG 0 1586 1603 5-8-5 1793 621865 101481 101498 GCTGGAGGGTTGGATCAG 16 1589 1606 5-8-5 1794 621866 101491 101508 ACACAGCAGGGCTGGAGG 34 1599 1616 5-8-5 1795 621867 101494 101511 GGCACACAGCAGGGCTGG 67 1602 1619 5-8-5 1796 621868 101499 101516 CTCTGGGCACACAGCAGG 75 1607 1624 5-8-5 1797 621869 101502 101519 TGGCTCTGGGCACACAGC 90 1610 1627 5-8-5 1798 621870 101505 101522 AGGTGGCTCTGGGCACAC 92 1613 1630 5-8-5 1799 621871 101508 101525 GGAAGGTGGCTCTGGGCA 67 1616 1633 5-8-5 1800 621872 101511 101528 AGAGGAAGGTGGCTCTGG 43 1619 1636 5-8-5 1801 621873 101514 101531 AGGAGAGGAAGGTGGCTC 65 1622 1639 5-8-5 1802 621874 101517 101534 TTTAGGAGAGGAAGGTGG 26 1625 1642 5-8-5 1803 621875 101520 101537 GTATTTAGGAGAGGAAGG 0 1628 1645 5-8-5 1804 621876 101537 101554 GAAGTGACAGAAGAGACG 37 1645 1662 5-8-5 1805 621877 101540 101557 CGGGAAGTGACAGAAGAG 59 1648 1665 5-8-5 1806 621878 101543 101560 GTTCGGGAAGTGACAGAA 65 1651 1668 5-8-5 1807 621879 101546 101563 CCAGTTCGGGAAGTGACA 35 1654 1671 5-8-5 1808 621880 101549 101566 CTGCCAGTTCGGGAAGTG 69 1657 1674 5-8-5 1809 621881 101552 101569 GAACTGCCAGTTCGGGAA 48 1660 1677 5-8-5 1810 621882 101555 101572 CCAGAACTGCCAGTTCGG 66 1663 1680 5-8-5 1811 621883 101558 101575 GCTCCAGAACTGCCAGTT 69 1666 1683 5-8-5 1812 621884 101561 101578 TTTGCTCCAGAACTGCCA 74 1669 1686 5-8-5 1813 621885 101564 101581 TCCTTTGCTCCAGAACTG 61 1672 1689 5-8-5 1814 621886 101567 101584 ATCTCCTTTGCTCCAGAA 64 1675 1692 5-8-5 1815 621887 101570 101587 TTCATCTCCTTTGCTCCA 36 1678 1695 5-8-5 1816 621888 101573 101590 AGTTTCATCTCCTTTGCT 32 1681 1698 5-8-5 1817

TABLE 35 Inhibition of tau mRNA by 5-10-5 MOE and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 and 2 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ISIS NO: 1 NO: 1 % NO: 2 NO: 2 ID NO start site stop site Sequence inhibition start site stop site Motif NO: 613412 73879 73898 GTGATCTTCCATCACTTCGA 87 345 364 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 95 n/a n/a 5-8-5 665 98928 98945 n/a n/a 621890 n/a n/a CCCTTGAGTTTCATCTCC 30 1687 1704 5-8-5 1818 621891 n/a n/a GCCCCCTTGAGTTTCATC 8 1690 1707 5-8-5 1819 621892 n/a n/a TCAGCCCCCTTGAGTTTC 0 1693 1710 5-8-5 1820 621893 n/a n/a CCATCAGCCCCCTTGAGT 4 1696 1713 5-8-5 1821 621894 n/a n/a TTACCATCAGCCCCCTTG 75 1699 1716 5-8-5 1822 621912 n/a n/a CGCAGAGCTGGGTGGTGT 0 1814 1831 5-8-5 1823 621913 n/a n/a AGTCGCAGAGCTGGGTGG 0 1817 1834 5-8-5 1824 621914 n/a n/a CTTAGTCGCAGAGCTGGG 21 1820 1837 5-8-5 1825 621915 n/a n/a TTGCTTAGTCGCAGAGCT 12 1823 1840 5-8-5 1826 621916 n/a n/a GACTTGCTTAGTCGCAGA 29 1826 1843 5-8-5 1827 621925 n/a n/a TTCACCTCTCTCAGATCT 4 1871 1888 5-8-5 1828 621926 n/a n/a AGGTTCACCTCTCTCAGA 9 1874 1891 5-8-5 1829 621927 n/a n/a TGGAGGTTCACCTCTCTC 7 1877 1894 5-8-5 1830 621928 n/a n/a TTTTGGAGGTTCACCTCT 11 1880 1897 5-8-5 1831 621889 101576 101593 TTGAGTTTCATCTCCTTT 23 1684 1701 5-8-5 1832 621895 102978 102995 GTTTTACCATCAGCCCCC 88 1702 1719 5-8-5 1833 621896 102995 103012 GCGGTGTGGCGATCTTCG 42 1719 1736 5-8-5 1834 621897 102998 103015 CCCGCGGTGTGGCGATCT 33 1722 1739 5-8-5 1835 621898 103001 103018 CTCCCCGCGGTGTGGCGA 37 1725 1742 5-8-5 1836 621899 103004 103021 CTGCTCCCCGCGGTGTGG 50 1728 1745 5-8-5 1837 621900 103007 103024 GGGCTGCTCCCCGCGGTG 43 1731 1748 5-8-5 1838 621901 103025 103042 GGCCCTTCTGGCCTGGAG 29 1749 1766 5-8-5 1839 621902 103029 103046 GCCTGGCCCTTCTGGCCT 34 1753 1770 5-8-5 1840 621903 103032 103049 TTGGCCTGGCCCTTCTGG 49 1756 1773 5-8-5 1841 621904 103050 103067 GCTGGAATCCTGGTGGCG 58 1774 1791 5-8-5 1842 621905 103053 103070 TTTGCTGGAATCCTGGTG 50 1777 1794 5-8-5 1843 621906 103056 103073 GTTTTTGCTGGAATCCTG 62 1780 1797 5-8-5 1844 621907 103075 103092 TGTCTTTGGAGCGGGCGG 21 1799 1816 5-8-5 1845 621908 103078 103095 TGGTGTCTTTGGAGCGGG 57 1802 1819 5-8-5 1846 621909 103081 103098 GGGTGGTGTCTTTGGAGC 17 1805 1822 5-8-5 1847 621910 103084 103101 GCTGGGTGGTGTCTTTGG 44 1808 1825 5-8-5 1848 621911 103087 103104 AGAGCTGGGTGGTGTCTT 47 1811 1828 5-8-5 1849 621917 105442 105459 CTGGACTTGCTTAGTCGC 27 1829 1846 5-8-5 1850 621918 105445 105462 TCTCTGGACTTGCTTAGT 0 1832 1849 5-8-5 1851 621919 105448 105465 TCTTCTCTGGACTTGCTT 48 1835 1852 5-8-5 1852 621920 105451 105468 TGGTCTTCTCTGGACTTG 49 1838 1855 5-8-5 1853 621921 105454 105471 GGGTGGTCTTCTCTGGAC 33 1841 1858 5-8-5 1854 621922 105473 105490 CAGATCTGGGCCCTGCAG 36 1860 1877 5-8-5 1855 621923 105476 105493 TCTCAGATCTGGGCCCTG 70 1863 1880 5-8-5 1856 621924 105479 105496 CTCTCTCAGATCTGGGCC 32 1866 1883 5-8-5 1857 621929 107917 107934 TGATTTTGGAGGTTCACC 44 1883 1900 5-8-5 1858 621930 107920 107937 CCCTGATTTTGGAGGTTC 74 1886 1903 5-8-5 1859 621931 107923 107940 ATCCCCTGATTTTGGAGG 26 1889 1906 5-8-5 1860 621932 107926 107943 GCGATCCCCTGATTTTGG 45 1892 1909 5-8-5 1861 621933 107929 107946 GCTGCGATCCCCTGATTT 38 1895 1912 5-8-5 1862 621934 107932 107949 GCCGCTGCGATCCCCTGA 42 1898 1915 5-8-5 1863 621935 107935 107952 GTAGCCGCTGCGATCCCC 31 1901 1918 5-8-5 1864 621936 107938 107955 GCTGTAGCCGCTGCGATC 67 1904 1921 5-8-5 1865 621937 107941 107958 GCTGCTGTAGCCGCTGCG 23 1907 1924 5-8-5 1866 621938 107971 107988 GCGGCTGCCGGGAGTGCC 40 1937 1954 5-8-5 1867 621939 107974 107991 GGAGCGGCTGCCGGGAGT 1 1940 1957 5-8-5 1868 621940 107978 107995 TGCGGGAGCGGCTGCCGG 48 1944 1961 5-8-5 1869 621941 108021 108038 ACCTTCTTGGGCTCCCGG 24 1987 2004 5-8-5 1870 621942 108024 108041 GCCACCTTCTTGGGCTCC 28 1990 2007 5-8-5 1871 621943 108027 108044 ACTGCCACCTTCTTGGGC 19 1993 2010 5-8-5 1872 621944 108030 108047 ACCACTGCCACCTTCTTG 28 1996 2013 5-8-5 1873 621945 108033 108050 CGGACCACTGCCACCTTC 72 1999 2016 5-8-5 1874 621946 108036 108053 GTACGGACCACTGCCACC 57 2002 2019 5-8-5 1875 621947 108039 108056 GGAGTACGGACCACTGCC 25 2005 2022 5-8-5 1876 621948 108042 108059 GGTGGAGTACGGACCACT 30 2008 2025 5-8-5 1877 621949 108045 108062 TTGGGTGGAGTACGGACC 24 2011 2028 5-8-5 1878 621950 108052 108069 CGGCGACTTGGGTGGAGT 1 2018 2035 5-8-5 1879 621951 108055 108072 AGACGGCGACTTGGGTGG 4 2021 2038 5-8-5 1880 621952 108058 108075 GGAAGACGGCGACTTGGG 50 2024 2041 5-8-5 1881 621953 108061 108078 GGCGGAAGACGGCGACTT 66 2027 2044 5-8-5 1882 621954 108064 108081 CTTGGCGGAAGACGGCGA 38 2030 2047 5-8-5 1883 621955 108067 108084 GCTCTTGGCGGAAGACGG 24 2033 2050 5-8-5 1884 621956 108070 108087 GCGGCTCTTGGCGGAAGA 20 2036 2053 5-8-5 1885 621957 108073 108090 CAGGCGGCTCTTGGCGGA 46 2039 2056 5-8-5 1886 621958 108076 108093 CTGCAGGCGGCTCTTGGC 72 2042 2059 5-8-5 1887 621959 108079 108096 TGTCTGCAGGCGGCTCTT 52 2045 2062 5-8-5 1888 621960 108082 108099 GGCTGTCTGCAGGCGGCT 47 2048 2065 5-8-5 1889 621961 108101 108118 GGTCTGGCATGGGCACGG 73 2067 2084 5-8-5 1890 621962 108137 108154 TCTCAGTGGAGCCGATCT 39 2103 2120 5-8-5 1891 621963 108140 108157 GGTTCTCAGTGGAGCCGA 66 2106 2123 5-8-5 1892 621964 108143 108160 TCAGGTTCTCAGTGGAGC 65 2109 2126 5-8-5 1893 621965 108146 108163 GCTTCAGGTTCTCAGTGG 44 2112 2129 5-8-5 1894

TABLE 36 Inhibition of tau mRNA by 5-10-5 MOE and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 and 2 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ISIS NO: 1 NO: 1 % NO: 2 NO: 2 ID NO start site stop site Sequence inhibition start site stop site Motif NO: 613412 73879 73898 GTGATCTTCCATCACTTCGA 88 345 364 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 94 n/a n/a 5-8-5 665 98928 98945 n/a n/a 621973 n/a n/a TGCACCTTCCCGCCTCCC 66 2137 2154 5-8-5 1895 621974 n/a n/a TATCTGCACCTTCCCGCC 17 2141 2158 5-8-5 1896 621975 n/a n/a AATTATCTGCACCTTCCC 27 2144 2161 5-8-5 1897 621994 n/a n/a TGCACACTGCCGCCTCCC 74 2230 2247 5-8-5 1898 621995 n/a n/a ACTATTTGCACACTGCCG 28 2236 2253 5-8-5 1899 621996 n/a n/a TAGACTATTTGCACACTG 29 2239 2256 5-8-5 1900 621997 n/a n/a TTGTAGACTATTTGCACA 62 2242 2259 5-8-5 1901 622012 n/a n/a CTCCTGGTTTATGATGGA 24 2310 2327 5-8-5 1902 622013 n/a n/a GGCCACCTCCTGGTTTAT 21 2316 2333 5-8-5 1903 622014 n/a n/a CACCTGGCCACCTCCTGG 44 2321 2338 5-8-5 1904 622041 n/a n/a ATCTTTTTATTTCCTCCG 32 2422 2439 5-8-5 1905 622042 n/a n/a TTTCAATCTTTTTATTTC 16 2427 2444 5-8-5 1906 621966 108149 108166 GGTGCTTCAGGTTCTCAG 48 2115 2132 5-8-5 1907 621967 108153 108170 GGCTGGTGCTTCAGGTTC 46 2119 2136 5-8-5 1908 621968 108156 108173 CCCGGCTGGTGCTTCAGG 50 2122 2139 5-8-5 1909 621969 108159 108176 CCTCCCGGCTGGTGCTTC 22 2125 2142 5-8-5 1910 621970 108162 108179 CCGCCTCCCGGCTGGTGC 27 2128 2145 5-8-5 1911 621971 108165 108182 TTCCCGCCTCCCGGCTGG 5 2131 2148 5-8-5 1912 621972 108168 108185 ACCTTCCCGCCTCCCGGC 39 2134 2151 5-8-5 1913 621976 121826 121843 ATTAATTATCTGCACCTT 76 2147 2164 5-8-5 1914 621977 121829 121846 CTTATTAATTATCTGCAC 73 2150 2167 5-8-5 1915 621978 121833 121850 GCTTCTTATTAATTATCT 68 2154 2171 5-8-5 1916 621979 121836 121853 CCAGCTTCTTATTAATTA 44 2157 2174 5-8-5 1917 621980 121839 121856 GATCCAGCTTCTTATTAA 31 2160 2177 5-8-5 1918 621981 121842 121859 TAAGATCCAGCTTCTTAT 44 2163 2180 5-8-5 1919 621982 121845 121862 TGCTAAGATCCAGCTTCT 69 2166 2183 5-8-5 1920 621983 121848 121865 CGTTGCTAAGATCCAGCT 82 2169 2186 5-8-5 1921 621984 121864 121881 CCACACTTGGACTGGACG 73 2185 2202 5-8-5 1922 621985 121867 121884 GAGCCACACTTGGACTGG 80 2188 2205 5-8-5 1923 621986 121870 121887 TTTGAGCCACACTTGGAC 59 2191 2208 5-8-5 1924 621987 121873 121890 TCCTTTGAGCCACACTTG 66 2194 2211 5-8-5 1925 621988 121876 121893 TTATCCTTTGAGCCACAC 77 2197 2214 5-8-5 1926 621989 121879 121896 ATATTATCCTTTGAGCCA 54 2200 2217 5-8-5 1927 621990 121882 121899 TTGATATTATCCTTTGAG 69 2203 2220 5-8-5 1928 621991 121885 121902 TGTTTGATATTATCCTTT 92 2206 2223 5-8-5 1929 621992 121903 121920 CTGCCGCCTCCCGGGACG 51 2224 2241 5-8-5 1930 621993 121906 121923 ACACTGCCGCCTCCCGGG 70 2227 2244 5-8-5 1931 621998 125764 125781 GGTTTGTAGACTATTTGC 76 2245 2262 5-8-5 1932 621999 125767 125784 ACTGGTTTGTAGACTATT 62 2248 2265 5-8-5 1933 622000 125770 125787 TCAACTGGTTTGTAGACT 39 2251 2268 5-8-5 1934 622001 125777 125794 GCTCAGGTCAACTGGTTT 87 2258 2275 5-8-5 1935 622002 125780 125797 CTTGCTCAGGTCAACTGG 53 2261 2278 5-8-5 1936 622003 125783 125800 CACCTTGCTCAGGTCAAC 68 2264 2281 5-8-5 1937 622004 125805 125822 CTAATGAGCCACACTTGG 51 2286 2303 5-8-5 1938 622005 125808 125825 TGCCTAATGAGCCACACT 65 2289 2306 5-8-5 1939 622006 125811 125828 TGTTGCCTAATGAGCCAC 69 2292 2309 5-8-5 1940 622007 125814 125831 GGATGTTGCCTAATGAGC 70 2295 2312 5-8-5 1941 622008 125817 125834 GATGGATGTTGCCTAATG 61 2298 2315 5-8-5 1942 622009 125820 125837 TATGATGGATGTTGCCTA 44 2301 2318 5-8-5 1943 622010 125823 125840 GTTTATGATGGATGTTGC 57 2304 2321 5-8-5 1944 622011 125826 125843 CTGGTTTATGATGGATGT 26 2307 2324 5-8-5 1945 622015 130141 130158 TTTACTTCCACCTGGCCA 36 2329 2346 5-8-5 1946 622016 130144 130161 GATTTTACTTCCACCTGG 49 2332 2349 5-8-5 1947 622017 130147 130164 TCAGATTTTACTTCCACC 76 2335 2352 5-8-5 1948 622018 130150 130167 TTCTCAGATTTTACTTCC 24 2338 2355 5-8-5 1949 622019 130153 130170 AGCTTCTCAGATTTTACT 30 2341 2358 5-8-5 1950 622020 130156 130173 TCAAGCTTCTCAGATTTT 39 2344 2361 5-8-5 1951 622021 130159 130176 AAGTCAAGCTTCTCAGAT 2 2347 2364 5-8-5 1952 622022 130162 130179 TTGAAGTCAAGCTTCTCA 53 2350 2367 5-8-5 1953 622023 130165 130182 TCCTTGAAGTCAAGCTTC 39 2353 2370 5-8-5 1954 622024 130168 130185 CTGTCCTTGAAGTCAAGC 19 2356 2373 5-8-5 1955 622025 130171 130188 ACTCTGTCCTTGAAGTCA 39 2359 2376 5-8-5 1956 622026 130174 130191 TGGACTCTGTCCTTGAAG 36 2362 2379 5-8-5 1957 622027 130177 130194 GACTGGACTCTGTCCTTG 75 2365 2382 5-8-5 1958 622028 130180 130197 TTCGACTGGACTCTGTCC 29 2368 2385 5-8-5 1959 622029 130183 130200 ATCTTCGACTGGACTCTG 33 2371 2388 5-8-5 1960 622030 130186 130203 CCAATCTTCGACTGGACT 49 2374 2391 5-8-5 1961 622031 130189 130206 GACCCAATCTTCGACTGG 74 2377 2394 5-8-5 1962 622032 130192 130209 AGGGACCCAATCTTCGAC 79 2380 2397 5-8-5 1963 622033 130195 130212 TCCAGGGACCCAATCTTC 77 2383 2400 5-8-5 1964 622034 130198 130215 TTGTCCAGGGACCCAATC 68 2386 2403 5-8-5 1965 622035 130201 130218 ATATTGTCCAGGGACCCA 6 2389 2406 5-8-5 1966 622036 130204 130221 GTGATATTGTCCAGGGAC 86 2392 2409 5-8-5 1967 622037 130207 130224 TGGGTGATATTGTCCAGG 68 2395 2412 5-8-5 1968 622038 130225 130242 TTTCCTCCGCCAGGGACG 77 2413 2430 5-8-5 1969 622039 130228 130245 TTATTTCCTCCGCCAGGG 52 2416 2433 5-8-5 1970 622040 130231 130248 TTTTTATTTCCTCCGCCA 61 2419 2436 5-8-5 1971

TABLE 37 Inhibition of tau mRNA by 5-10-5 MOE and 5-8-5 MOE gapmers targeting SEQ ID NO: 1 and 2 SEQ ID SEQ ID SEQ ID SEQ ID ISIS NO: 1 NO: 1 % NO: 2 NO: 2 SEQ NO start site stop site Sequence inhibition start site stop site Motif ID NO: 613412 73879 73898 GTGATCTTCCATCACTTCGA 89 345 364 5-10-5 25 620887 98891 98908 GTTTTCAAACACACCTTC 98 n/a n/a 5-8-5 665 98928 98945 n/a n/a 622043 n/a n/a GGGTTTCAATCTTTTTAT 65 2430 2447 5-8-5 1972 622044 135477 135494 GTCAGCTTGTGGGTTTCA 86 2440 2457 5-8-5 1973 622045 135480 135497 AAGGTCAGCTTGTGGGTT 49 2443 2460 5-8-5 1974 622046 135484 135501 GCGGAAGGTCAGCTTGTG 26 2447 2464 5-8-5 1975 622047 135488 135505 TCTCGCGGAAGGTCAGCT 54 2451 2468 5-8-5 1976 622048 135491 135508 CGTTCTCGCGGAAGGTCA 63 2454 2471 5-8-5 1977 622049 135507 135524 TCTGTCTTGGCTTTGGCG 30 2470 2487 5-8-5 1978 622050 135510 135527 TGGTCTGTCTTGGCTTTG 72 2473 2490 5-8-5 1979 622051 135513 135530 CCGTGGTCTGTCTTGGCT 75 2476 2493 5-8-5 1980 622052 135516 135533 GCCCCGTGGTCTGTCTTG 39 2479 2496 5-8-5 1981 622053 135527 135544 ACACGATCTCCGCCCCGT 80 2490 2507 5-8-5 1982 622054 135530 135547 TGTACACGATCTCCGCCC 72 2493 2510 5-8-5 1983 622055 135533 135550 ACTTGTACACGATCTCCG 30 2496 2513 5-8-5 1984 622056 135536 135553 GCGACTTGTACACGATCT 72 2499 2516 5-8-5 1985 622057 135539 135556 CTGGCGACTTGTACACGA 65 2502 2519 5-8-5 1986 622058 135542 135559 CCACTGGCGACTTGTACA 35 2505 2522 5-8-5 1987 622059 135545 135562 ACACCACTGGCGACTTGT 46 2508 2525 5-8-5 1988 622060 135548 135565 CAGACACCACTGGCGACT 61 2511 2528 5-8-5 1989 622061 135551 135568 CCCCAGACACCACTGGCG 74 2514 2531 5-8-5 1990 622062 135554 135571 TGTCCCCAGACACCACTG 38 2517 2534 5-8-5 1991 622063 135572 135589 TGAGATGCCGTGGAGACG 36 2535 2552 5-8-5 1992 622064 135575 135592 TGCTGAGATGCCGTGGAG 34 2538 2555 5-8-5 1993 622065 135599 135616 CGATGCTGCCGGTGGAGG 43 2562 2579 5-8-5 1994 622066 135602 135619 TGTCGATGCTGCCGGTGG 51 2565 2582 5-8-5 1995 622067 135605 135622 CCATGTCGATGCTGCCGG 66 2568 2585 5-8-5 1996 622068 135608 135625 CTACCATGTCGATGCTGC 68 2571 2588 5-8-5 1997 622069 135611 135628 AGTCTACCATGTCGATGC 56 2574 2591 5-8-5 1998 622070 135614 135631 GCGAGTCTACCATGTCGA 68 2577 2594 5-8-5 1999 622071 135654 135671 AGGGAGGCAGACACCTCG 24 2617 2634 5-8-5 2000 622072 135657 135674 GCCAGGGAGGCAGACACC 73 2620 2637 5-8-5 2001 622073 135661 135678 CTTGGCCAGGGAGGCAGA 50 2624 2641 5-8-5 2002 622074 135664 135681 CTGCTTGGCCAGGGAGGC 44 2627 2644 5-8-5 2003 622075 135667 135684 ACCCTGCTTGGCCAGGGA 18 2630 2647 5-8-5 2004 622076 135678 135695 CCTGATCACAAACCCTGC 65 2641 2658 5-8-5 2005 622077 135681 135698 GGGCCTGATCACAAACCC 82 2644 2661 5-8-5 2006 622078 135697 135714 TTATTGACCGCCCCAGGG 42 2660 2677 5-8-5 2007 622079 135700 135717 CAATTATTGACCGCCCCA 54 2663 2680 5-8-5 2008 622080 135703 135720 CCACAATTATTGACCGCC 95 2666 2683 5-8-5 2009 622081 135706 135723 TCTCCACAATTATTGACC 63 2669 2686 5-8-5 2010 622082 135709 135726 TCCTCTCCACAATTATTG 41 2672 2689 5-8-5 2011 622083 135712 135729 CTCTCCTCTCCACAATTA 76 2675 2692 5-8-5 2012 622084 135715 135732 ATTCTCTCCTCTCCACAA 57 2678 2695 5-8-5 2013 622085 135718 135735 CTCATTCTCTCCTCTCCA 72 2681 2698 5-8-5 2014 622086 135721 135738 TCTCTCATTCTCTCCTCT 69 2684 2701 5-8-5 2015 622087 135724 135741 CACTCTCTCATTCTCTCC 75 2687 2704 5-8-5 2016 622088 135727 135744 CCACACTCTCTCATTCTC 90 2690 2707 5-8-5 2017 622089 135730 135747 TTTCCACACTCTCTCATT 70 2693 2710 5-8-5 2018 622090 135733 135750 TTTTTTCCACACTCTCTC 76 2696 2713 5-8-5 2019 622091 135737 135754 CTTTTTTTTTCCACACTC 84 2700 2717 5-8-5 2020 622092 135740 135757 ATTCTTTTTTTTTCCACA 67 2703 2720 5-8-5 2021 622093 135746 135763 GTCATTATTCTTTTTTTT 54 2709 2726 5-8-5 2022 622094 135749 135766 CGGGTCATTATTCTTTTT 82 2712 2729 5-8-5 2023 622095 135752 135769 GGCCGGGTCATTATTCTT 72 2715 2732 5-8-5 2024 622096 135783 135800 CTGCGAGGAGCAGCTGGG 52 2746 2763 5-8-5 2025 622097 135786 135803 GAACTGCGAGGAGCAGCT 59 2749 2766 5-8-5 2026 622098 135789 135806 ACCGAACTGCGAGGAGCA 77 2752 2769 5-8-5 2027 622099 135792 135809 TTAACCGAACTGCGAGGA 62 2755 2772 5-8-5 2028 622100 135795 135812 CAATTAACCGAACTGCGA 55 2758 2775 5-8-5 2029 622101 135798 135815 AACCAATTAACCGAACTG 59 2761 2778 5-8-5 2030 622102 135801 135818 ATTAACCAATTAACCGAA 71 2764 2781 5-8-5 2031 622103 135804 135821 GTGATTAACCAATTAACC 70 2767 2784 5-8-5 2032 622104 135807 135824 TAAGTGATTAACCAATTA 42 2770 2787 5-8-5 2033 622105 135810 135827 GGTTAAGTGATTAACCAA 42 2773 2790 5-8-5 2034 622106 135813 135830 GCAGGTTAAGTGATTAAC 78 2776 2793 5-8-5 2035 622107 135816 135833 AAAGCAGGTTAAGTGATT 38 2779 2796 5-8-5 2036 622108 135819 135836 ACAAAAGCAGGTTAAGTG 55 2782 2799 5-8-5 2037 622109 135822 135839 GTGACAAAAGCAGGTTAA 82 2785 2802 5-8-5 2038 622110 135825 135842 CGAGTGACAAAAGCAGGT 86 2788 2805 5-8-5 2039 622111 135828 135845 AGCCGAGTGACAAAAGCA 93 2791 2808 5-8-5 2040 622112 135831 135848 CAAAGCCGAGTGACAAAA 73 2794 2811 5-8-5 2041 622113 135834 135851 AGCCAAAGCCGAGTGACA 82 2797 2814 5-8-5 2042 622114 135837 135854 CCGAGCCAAAGCCGAGTG 82 2800 2817 5-8-5 2043 622115 135840 135857 GTCCCGAGCCAAAGCCGA 63 2803 2820 5-8-5 2044 622116 135843 135860 GAAGTCCCGAGCCAAAGC 59 2806 2823 5-8-5 2045 622117 135846 135863 TTTGAAGTCCCGAGCCAA 76 2809 2826 5-8-5 2046 622118 135849 135866 GATTTTGAAGTCCCGAGC 72 2812 2829 5-8-5 2047 622119 135852 135869 ACTGATTTTGAAGTCCCG 73 2815 2832 5-8-5 2048

TABLE 38 Inhibition of tau mRNA by 5-8-5 MOE gapmers targeting SEQ ID NO: 1 SEQ ID SEQ ID SEQ NO: 1 start NO: 1 % ID ISIS NO site stop site Sequence inhibition Motif NO: 620887 98891 98908 GTTTTCAAACACACCTTC 96 5-8-5 665 98928 98945 623853 12069 12086 TGTTTGGATTTCTATCGG 82 5-8-5 2049 623854 12077 12094 TGACATGGTGTTTGGATT 47 5-8-5 2050 623855 12079 12096 GCTGACATGGTGTTTGGA 77 5-8-5 2051 623856 12080 12097 CGCTGACATGGTGTTTGG 78 5-8-5 2052 623857 12081 12098 TCGCTGACATGGTGTTTG 83 5-8-5 2053 623858 12082 12099 CTCGCTGACATGGTGTTT 84 5-8-5 2054 623859 12083 12100 ACTCGCTGACATGGTGTT 74 5-8-5 2055 623860 12084 12101 GACTCGCTGACATGGTGT 89 5-8-5 2056 623861 12085 12102 GGACTCGCTGACATGGTG 77 5-8-5 2057 623862 16753 16770 TTATCCAGGAAAAATATT 21 5-8-5 2058 623863 16756 16773 GGATTATCCAGGAAAAAT 86 5-8-5 2059 623864 16758 16775 TAGGATTATCCAGGAAAA 76 5-8-5 2060 623865 16759 16776 ATAGGATTATCCAGGAAA 84 5-8-5 2061 623866 16760 16777 TATAGGATTATCCAGGAA 62 5-8-5 2062 623867 16761 16778 CTATAGGATTATCCAGGA 87 5-8-5 2063 623868 16763 16780 CCCTATAGGATTATCCAG 76 5-8-5 2064 623869 16764 16781 TCCCTATAGGATTATCCA 34 5-8-5 2065 623870 16766 16783 TATCCCTATAGGATTATC 56 5-8-5 2066 623871 16769 16786 AGTTATCCCTATAGGATT 16 5-8-5 2067 623872 16774 16791 AGGCAAGTTATCCCTATA 86 5-8-5 2068 623873 17416 17433 ATCAGCACCTGACTGCGG 78 5-8-5 2069 623874 17421 17438 TTCAGATCAGCACCTGAC 91 5-8-5 2070 623875 17424 17441 GACTTCAGATCAGCACCT 92 5-8-5 2071 623876 17426 17443 AAGACTTCAGATCAGCAC 92 5-8-5 2072 623877 17427 17444 AAAGACTTCAGATCAGCA 95 5-8-5 2073 623878 17428 17445 CAAAGACTTCAGATCAGC 96 5-8-5 2074 623879 17429 17446 CCAAAGACTTCAGATCAG 86 5-8-5 2075 623880 17430 17447 ACCAAAGACTTCAGATCA 65 5-8-5 2076 623881 17431 17448 CACCAAAGACTTCAGATC 51 5-8-5 2077 623882 17432 17449 CCACCAAAGACTTCAGAT 55 5-8-5 2078 623883 17434 17451 GCCCACCAAAGACTTCAG 77 5-8-5 2079 623884 17437 17454 TCAGCCCACCAAAGACTT 56 5-8-5 2080 623885 17442 17459 TAAAGTCAGCCCACCAAA 2 5-8-5 2081 623886 20943 20960 GTTATTGGGACTGACCTT 69 5-8-5 2082 623887 20948 20965 GATTTGTTATTGGGACTG 78 5-8-5 2083 613412 73879 73898 GTGATCTTCCATCACTTCGA 89 5-8-5 25 623811 122370 122387 CAGTGGAGCCACTCAACG 41 5-8-5 2084 623812 122380 122397 CACCTGTCCACAGTGGAG 58 5-8-5 2085 623813 122391 122408 AACAAACGGGTCACCTGT 63 5-8-5 2086 623814 122445 122462 CGTGTAGGAGCAGCAGCT 81 5-8-5 2087 623815 122538 122555 CTTTGGTTTGGCTCTTTG 64 5-8-5 2088 623816 123104 123121 GCTGGTGGGAGAGGAGCC 9 5-8-5 2089 623817 123288 123305 ATGCGGGTGGCTGCCTCA 65 5-8-5 2090 623818 123293 123310 GCTGGATGCGGGTGGCTG 24 5-8-5 2091 623819 123341 123358 GTGCTCAGGGCAGGAAGC 45 5-8-5 2092 623820 123590 123607 CAAAGCTCAAACAGCTGA 56 5-8-5 2093 623821 123601 123618 AGAACCAGGATCAAAGCT 41 5-8-5 2094 623822 123704 123721 GCTCCGCCTCAGCAGCAC 71 5-8-5 2095 623823 123722 123739 GTGTGCACTCTCTCCCCA 45 5-8-5 2096 623824 123877 123894 GGCTAGCCCGCAGACGAG 31 5-8-5 2097 623825 123976 123993 CCTGTGAAGGTGCTCAGA 58 5-8-5 2098 623826 124033 124050 GATCCAGTGCCCCCAGAT 54 5-8-5 2099 623827 124057 124074 GGAGAGGCTAGGGCTCAG 68 5-8-5 2100 623828 124223 124240 AGAGGAGTGGATGGCAGT 37 5-8-5 2101 623829 124293 124310 TGGCCTGGGTGAGGTAAC 23 5-8-5 2102 623830 124296 124313 CTCTGGCCTGGGTGAGGT 28 5-8-5 2103 623831 124596 124613 AGGGCAGGTGGTGGTTTC 8 5-8-5 2104 623832 124780 124797 GACATTCCTGGAGTCCCC 83 5-8-5 2105 623833 124859 124876 TGGCAGACAGACAGGTCC 87 5-8-5 2106 623834 124909 124926 AGTCAAAGAACCAGCTCC 68 5-8-5 2107 623835 124949 124966 TCCCTCTGGGAATGATGA 41 5-8-5 2108 623836 124965 124982 GCCTCCAGGGCACCGCTC 76 5-8-5 2109 623837 124972 124989 GCCTGTGGCCTCCAGGGC 43 5-8-5 2110 623838 124977 124994 AGGAGGCCTGTGGCCTCC 11 5-8-5 2111 623839 125075 125092 AATAGGAACAAAGCAACA 25 5-8-5 2112 623840 125086 125103 CTGTCTTTAGCAATAGGA 84 5-8-5 2113 623841 125100 125117 TGTCCTGGACATTCCTGT 66 5-8-5 2114 623842 125291 125308 TGGAAAGGCAGGAGTGGG 0 5-8-5 2115 623843 125306 125323 TCTGAAAAATCTTGCTGG 51 5-8-5 2116 623844 125310 125327 AGCATCTGAAAAATCTTG 82 5-8-5 2117 623845 125322 125339 ATGAGTATGCACAGCATC 68 5-8-5 2118 623846 125337 125354 AAAGTGATCAATATGATG 33 5-8-5 2119 623847 125390 125407 GTCACTCCCTTTCCTGAC 68 5-8-5 2120 623848 125408 125425 ACGCTTAAGTGTAAAAAT 38 5-8-5 2121 623849 125557 125574 AAATGTGTTGTCGAAATT 19 5-8-5 2122 623850 125567 125584 CAGGGTGGAAAAATGTGT 33 5-8-5 2123 623851 125598 125615 TTCCCAGCTGCCATGAGG 33 5-8-5 2124 623852 125743 125760 TGGAGATGAGAGAGGAGG 0 5-8-5 2125

TABLE 39 Inhibition of Tau mRNA by 5-8-5 MOE gapmers targeting SEQ ID NO: 1 SEQ ID SEQ ID SEQ NO: 1 start NO: 1 stop % ID ISIS NO site site Sequence inhibition Motif NO: 623661 6196 6213 GCGCTTACCTGATAGTCG 36 5-8-5 2126 613412 73879 73898 GTGATCTTCCATCACTTCGA 87 5-8-5 25 623662 73981 73998 TCCACTAACCTTTCAGGC 30 5-8-5 2127 623663 83368 83385 GGGAGATTCTGGAACACA 72 5-8-5 2128 623664 83455 83472 GGGCCCACCTTCCGCTGT 22 5-8-5 2129 623665 85895 85912 CTGTCACATCTAGAAACC 21 5-8-5 2130 623666 85982 85999 TACCCTCACCTGTGGTTC 0 5-8-5 2131 623667 89885 89902 CTTCTTCAGCTGGTGTAT 22 5-8-5 2132 623668 89951 89968 TTCACTGACCTTGGGTCA 0 5-8-5 2133 623669 94688 94705 TTTCAGGCTCTGTGTGGA 61 5-8-5 2134 623670 95441 95458 GACACAGACCTTTGAGTT 53 5-8-5 2135 623671 98550 98567 CCATGCGAGCTGATAAAA 39 5-8-5 2136 621344 98607 98624 GTCAGCTTACCTTGGCTT 34 5-8-5 814 620887 98891 98908 GTTTTCAAACACACCTTC 94 5-8-5 665 98928 98945 623672 101387 101404 GTGGATGTCTTAAACATA 89 5-8-5 2137 623673 101586 101603 TTTCCTTACCTTGAGTTT 0 5-8-5 2138 623674 102970 102987 ATCAGCCCCCTGTAAATG 7 5-8-5 2139 623675 103097 103114 TCTTCTTACCAGAGCTGG 0 5-8-5 2140 623676 107909 107926 GAGGTTCACCTGGGAAGG 0 5-8-5 2141 623677 108175 108192 CACTCTCACCTTCCCGCC 6 5-8-5 2142 623712 108255 108272 CCTCCAGGCGCAGCCCTA 92 5-8-5 2143 623713 108413 108430 CCTGAGGAGGGCACTCAC 28 5-8-5 2144 623714 108530 108547 GGAACCGCTGTGGGTGCC 60 5-8-5 2145 623715 108565 108582 TGGGTGGCAGTGTATTCT 63 5-8-5 2146 623716 108690 108707 GGAGAGCTCGCGAGCACC 69 5-8-5 2147 623717 108896 108913 GAGGTGGCTACCCACGGC 59 5-8-5 2148 623718 109561 109578 CCAGAAGGCCCAGCACAT 51 5-8-5 2149 623719 109574 109591 CGAGGCCCAGTGCCCAGA 59 5-8-5 2150 623720 109607 109624 AGGCCCAGGGTTCCAGAA 55 5-8-5 2151 623721 109623 109640 AGGCAAGCTGACACGCAG 73 5-8-5 2152 623722 110101 110118 CAGGAAAAGGCCGGACCT 37 5-8-5 2153 623723 110103 110120 AGCAGGAAAAGGCCGGAC 78 5-8-5 2154 623724 110182 110199 TTCCCCAAGGTCTCTAAC 14 5-8-5 2155 623725 110413 110430 AGGAAAGGCCAGTGAGGG 8 5-8-5 2156 623726 110500 110517 TTGTTATGTGACTTGAGG 74 5-8-5 2157 623727 110630 110647 ATTCCCCACCATGGGACA 17 5-8-5 2158 623728 110635 110652 AGGACATTCCCCACCATG 9 5-8-5 2159 623729 110648 110665 AGATAAGGAGAGAAGGAC 15 5-8-5 2160 623730 110755 110772 ATGCTCAGTGTGGTCAGA 73 5-8-5 2161 623731 110852 110869 AGGGCCGGCCACCTGCAC 56 5-8-5 2162 623732 110919 110936 TCTGTCTCTGGCAACCTG 85 5-8-5 2163 623733 110954 110971 AACAGGGAAGCTACTTCC 6 5-8-5 2164 623734 111077 111094 GGGCCTTCAATGGAAAGT 21 5-8-5 2165 623735 111153 111170 GGTCGCCTGACTCTCACC 66 5-8-5 2166 623736 111174 111191 CCCTTTCTACACTTGGCA 81 5-8-5 2167 623678 121820 121837 TATCTGCACCTTTGGTAG 30 5-8-5 2168 623685 121830 121847 TCTTATTAATTATCTGCA 81 5-8-5 2169 623686 121831 121848 TTCTTATTAATTATCTGC 74 5-8-5 2170 623687 121834 121851 AGCTTCTTATTAATTATC 46 5-8-5 2171 623688 121835 121852 CAGCTTCTTATTAATTAT 6 5-8-5 2172 623689 121837 121854 TCCAGCTTCTTATTAATT 4 5-8-5 2173 623690 121838 121855 ATCCAGCTTCTTATTAAT 2 5-8-5 2174 623691 121840 121857 AGATCCAGCTTCTTATTA 2 5-8-5 2175 623692 121841 121858 AAGATCCAGCTTCTTATT 17 5-8-5 2176 623693 121843 121860 CTAAGATCCAGCTTCTTA 41 5-8-5 2177 623694 121844 121861 GCTAAGATCCAGCTTCTT 75 5-8-5 2178 623695 121846 121863 TTGCTAAGATCCAGCTTC 50 5-8-5 2179 623696 121847 121864 GTTGCTAAGATCCAGCTT 52 5-8-5 2180 623697 121865 121882 GCCACACTTGGACTGGAC 88 5-8-5 2181 623698 121866 121883 AGCCACACTTGGACTGGA 89 5-8-5 2182 623699 121868 121885 TGAGCCACACTTGGACTG 59 5-8-5 2183 623700 121869 121886 TTGAGCCACACTTGGACT 62 5-8-5 2184 623701 121872 121889 CCTTTGAGCCACACTTGG 69 5-8-5 2185 623702 121874 121891 ATCCTTTGAGCCACACTT 64 5-8-5 2186 623703 121875 121892 TATCCTTTGAGCCACACT 69 5-8-5 2187 623704 121877 121894 ATTATCCTTTGAGCCACA 76 5-8-5 2188 623705 121878 121895 TATTATCCTTTGAGCCAC 76 5-8-5 2189 623706 121880 121897 GATATTATCCTTTGAGCC 54 5-8-5 2190 623707 121881 121898 TGATATTATCCTTTGAGC 65 5-8-5 2191 623708 121884 121901 GTTTGATATTATCCTTTG 94 5-8-5 2192 623709 121886 121903 GTGTTTGATATTATCCTT 85 5-8-5 2193 623710 121887 121904 CGTGTTTGATATTATCCT 78 5-8-5 2194 623711 121904 121921 ACTGCCGCCTCCCGGGAC 50 5-8-5 2195 623679 121913 121930 GGTACTCACACTGCCGCC 72 5-8-5 1654 623680 125753 125770 TATTTGCACCTGGAGATG 0 5-8-5 2196 623681 125835 125852 CAGGGCTACCTGGTTTAT 26 5-8-5 2197 623682 130128 130145 GGCCACCTCCTAGAACAC 8 5-8-5 2198 623683 130241 130258 CCCCTTTACCTTTTTATT 0 5-8-5 2199 623684 135466 135483 GGTTTCAATCTGCAAGAA 39 5-8-5 2200

Example 11 Dose-Dependent Antisense Inhibition of Human Tau in SH-SY5Y Cells

Gapmers from studies described above exhibiting significant in vitro inhibition of tau mRNA were selected and tested at various doses in SH-SY5Y cells. The antisense oligonucleotides were tested in a series of experiments that had similar culture conditions. The results for each experiment are presented in separate tables shown below. Cells were plated at a density of 20,000 cells per well and transfected using electroporation with 0.938 μM, 0.1.875 μM, 3.750 μM, 7.500 μM, and 15.00 μM concentrations of antisense oligonucleotide, as specified in the Tables below. After a treatment period of approximately 16 hours, RNA was isolated from the cells and tau mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3104 was used to measure mRNA levels. Tau mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of tau, relative to untreated control cells. Tau mRNA levels were significantly reduced in a dose-dependent manner in antisense oligonucleotide treated cells.

TABLE 40 ISIS No 0.938 μM 1.875 μM 3.750 μM 7.500 μM 15.00 μM 613369 57 67 81 91 96 613370 83 91 96 98 97 613371 68 78 95 95 97 613412 41 55 73 86 96 620887 53 76 90 93 95 621238 58 80 76 93 96 621251 68 77 87 89 90 621263 57 69 81 92 95 621302 46 66 80 78 87 621309 56 64 84 86 90 621311 38 54 69 77 86 621312 42 36 57 83 90 621318 40 54 71 87 87 621346 42 46 60 64 78 621870 42 62 76 85 92

TABLE 41 ISIS No 0.938 μM 1.875 μM 3.750 μM 7.500 μM 15.00 μM 613412 14 45 68 84 92 620887 57 74 87 93 95 620888 63 78 76 94 96 620890 76 86 92 96 97 620891 67 79 91 96 96 620918 38 55 65 81 88 620940 38 52 73 89 96 620947 26 47 58 78 87 620958 32 38 61 79 93 621013 49 75 86 92 94 621049 38 45 68 78 80 621056 40 45 70 81 90 621078 23 28 48 67 84 621082 22 32 53 62 90 621088 26 43 59 80 92

TABLE 42 ISIS No 0.938 μM 1.875 μM 3.750 μM 7.500 μM 15.00 μM 613412 17 45 62 84 89 620887 52 71 91 96 95 621147 28 37 59 69 84 621181 39 61 78 85 83 621183 31 43 64 72 87 621836 24 39 64 81 93 621838 33 59 70 91 97 621844 20 38 64 80 91 621845 30 37 62 72 90 621861 19 35 59 80 92 621869 30 56 64 83 91 622125 14 28 52 73 85 622129 26 36 60 71 84 622170 18 42 72 77 91 622196 11 39 59 74 89

TABLE 43 ISIS No 0.938 μM 1.875 μM 3.750 μM 7.500 μM 15.00 μM 613412 21 42 64 82 95 620887 60 79 92 95 95 623746 19 43 54 81 91 623750 50 63 75 83 90 623758 37 54 72 80 82 623805 25 33 48 74 86 623807 15 29 44 75 88 623833 16 60 65 86 89 623860 51 71 80 85 85 623867 9 51 48 74 86 623874 24 28 72 84 94 623875 38 62 80 93 95 623876 40 63 83 93 95 623877 59 76 92 97 96 623878 50 73 90 94 95

TABLE 44 ISIS No 0.938 μM 1.875 μM 3.750 μM 7.500 μM 15.00 μM 613412 8 25 58 77 87 620887 49 77 87 95 86 621360 27 46 68 83 90 621361 52 73 89 92 94 621363 28 45 71 76 77 621364 39 58 77 90 95 621407 38 60 75 82 78 621414 41 66 80 90 83 621424 54 72 87 94 90 621425 8 22 52 74 84 621426 33 46 63 84 91 621740 0 1 6 33 65 621793 4 15 30 41 52 621794 29 38 57 78 85 621810 15 39 45 73 79

TABLE 45 ISIS No 0.938 μM 1.875 μM 3.750 μM 7.500 μM 15.00 μM 613412 40 72 78 92 94 620887 58 79 92 96 97 621431 62 63 79 85 87 621441 71 80 84 95 95 621446 53 70 82 89 94 621448 46 60 85 89 93 621454 63 70 81 89 94 621484 60 65 75 88 93 621570 26 55 81 79 90 621576 23 34 71 75 80 621578 30 45 67 81 90 621579 14 28 49 69 88 621598 38 55 73 88 94 621670 37 57 79 82 92 621675 31 35 51 79 87

TABLE 46 ISIS No 0.938 μM 1.875 μM 3.750 μM 7.500 μM 15.00 μM 613412 21 38 70 84 92 620887 50 63 83 91 96 621894 35 16 32 54 70 621895 23 48 61 82 89 621961 8 14 47 57 75 621983 33 52 63 72 83 621991 36 56 68 86 92 622001 8 33 60 77 90 622080 29 53 69 67 93 622088 20 45 66 84 91 622091 14 33 47 70 82 622110 25 24 64 80 90 622111 17 41 74 86 92 623747 12 47 53 82 87

TABLE 47 ISIS No 0.938 μM 1.875 μM 3.750 μM 7.500 μM 15.00 μM 613412 22 34 49 77 77 620887 51 68 83 94 95 623672 33 43 64 82 88 623697 37 53 59 80 90 623698 32 57 63 84 92 623708 47 68 82 88 95 623709 14 37 62 76 89 623712 22 45 70 86 92 623732 18 37 54 76 87 623756 23 38 68 86 94 623782 50 72 83 90 93 623791 58 71 87 93 95 623796 44 57 74 79 85 623808 30 55 73 84 91 623809 31 42 66 82 93

Example 12 Antisense Inhibition of Human Tau in HepG2 Cells by 5-10-5 MOE, 5-8-5 MOE, 4-8-6 MOE, or 6-8-4 MOE Gapmers

Antisense oligonucleotides were designed targeting a tau nucleic acid and were tested for their effects on tau mRNA in vitro. The antisense oligonucleotides were tested in a series of experiments that had similar culture conditions. ISIS 613412 was also included in the assays. The results for each experiment are presented in separate tables shown below. Cultured HepG2 cells at a density of 20,000 cells per well were transfected using electroporation with 8,000 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and tau mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3104 was used to measure mRNA levels. Tau mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of tau, relative to untreated control cells.

The newly designed chimeric antisense oligonucleotides in the Tables below were designed as 5-8-5 MOE, 4-8-6 MOE, or 6-8-4 MOE gapmers. The 5-8-5 MOE gapmers are 18 nucleosides in length, wherein the central gap segment comprises of eight 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five nucleosides each. The 4-8-6 MOE gapmers are 18 nucleosides in length, wherein the central gap segment comprises of eight 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising four and six nucleosides respectively. The 6-8-4 MOE gapmers are 18 nucleosides in length, wherein the central gap segment comprises of eight 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising six and four nucleosides respectively. Each nucleoside in the 5′ wing segment and each nucleoside in the 3′ wing segment has a 2′-MOE modification. The internucleoside linkage motif throughout for each gapmer in the tables below, except for ISIS 613412, is 5′-sooosssssssssooss-3′, wherein each “s” represents a phosphorothioate internucleoside linkage and wherein each “o” represents a phosphodiester internucleoside linkage. The internucleoside linkage motif for ISIS 613412 is 5′-soooossssssssssooss-3′, wherein each “s” represents a phosphorothioate internucleoside linkage and wherein each “o” represents a phosphodiester internucleoside linkage. All cytosine residues throughout each gapmer are 5-methylcytosines. “Start site” indicates the 5′-most nucleoside to which the gapmer is targeted in the human gene sequence. “Stop site” indicates the 3′-most nucleoside to which the gapmer is targeted human gene sequence. Each gapmer listed in the Tables below is targeted to human tau genomic sequence, designated herein as SEQ ID NO: 1 (GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to 9381000).

TABLE 48 Inhibition of tau mRNA by 5-10-5 MOE gapmers targeting SEQ ID NO: 1 SEQ SEQ ID ID NO: 1 NO: 1 SEQ ISIS Start Stop % ID NO Site Site Sequence Motif inhibition NO 613412 73879 73898 GTGATCTTCCATCACTTCGA 5-10-5 90 25 620887 98891 98908 GTTTTCAAACACACCTTC 5-10-5 93 665 98928 98945 623888 20951 20968 TTCGATTTGTTATTGGGA 5-10-5 79 2201 623889 20953 20970 ACTTCGATTTGTTATTGG 5-10-5 61 2202 623890 20954 20971 GACTTCGATTTGTTATTG 5-10-5 30 2203 623891 20955 20972 TGACTTCGATTTGTTATT 5-10-5 62 2204 623892 20956 20973 CTGACTTCGATTTGTTAT 5-10-5 60 2205 623893 20957 20974 GCTGACTTCGATTTGTTA 5-10-5 43 2206 623894 20958 20975 AGCTGACTTCGATTTGTT 5-10-5 83 2207 623895 20959 20976 CAGCTGACTTCGATTTGT 5-10-5 68 2208 623896 20961 20978 CCCAGCTGACTTCGATTT 5-10-5 11 2209 623897 20964 20981 ACGCCCAGCTGACTTCGA 5-10-5 65 2210 623898 27148 27165 TGCCTTATATATGCTGAA 5-10-5 82 2211 623899 27153 27170 TTACATGCCTTATATATG 5-10-5 31 2212 623900 27156 27173 CAGTTACATGCCTTATAT 5-10-5 33 2213 623901 27158 27175 TTCAGTTACATGCCTTAT 5-10-5 72 2214 623902 27159 27176 GTTCAGTTACATGCCTTA 5-10-5 92 2215 623903 27169 27186 AAAGTGCTGTGTTCAGTT 5-10-5 71 2216 623904 27174 27191 CCTCTAAAGTGCTGTGTT 5-10-5 35 2217 623905 28215 28232 GACTTTTCTCAATGTAAC 5-10-5 43 2218 623906 28220 28237 CTGCAGACTTTTCTCAAT 5-10-5 61 2219 623907 28226 28243 ACCTCTCTGCAGACTTTT 5-10-5 79 2220 623908 28227 28244 CACCTCTCTGCAGACTTT 5-10-5 81 2221 623909 28228 28245 GCACCTCTCTGCAGACTT 5-10-5 82 2222 623910 28229 28246 GGCACCTCTCTGCAGACT 5-10-5 81 2223 623911 28230 28247 TGGCACCTCTCTGCAGAC 5-10-5 36 2224 623912 28231 28248 CTGGCACCTCTCTGCAGA 5-10-5 11 2225 623913 28233 28250 TGCTGGCACCTCTCTGCA 5-10-5 51 2226 623914 28236 28253 TGATGCTGGCACCTCTCT 5-10-5 63 2227 623915 28241 28258 CCTTGTGATGCTGGCACC 5-10-5 79 2228 623916 75133 75150 TTTTTAGCATTAAAAGAG 5-10-5 0 2229 623917 75144 75161 GTGTTTTCTTATTTTTAG 5-10-5 80 2230 623918 75149 75166 GCAAGGTGTTTTCTTATT 5-10-5 55 2231 623919 86762 86779 TAGGCCTCTTCTGCATTT 5-10-5 27 2232 623920 86767 86784 CTGGCTAGGCCTCTTCTG 5-10-5 54 2233 623921 86770 86787 ATCCTGGCTAGGCCTCTT 5-10-5 69 2234 623922 86772 86789 AAATCCTGGCTAGGCCTC 5-10-5 67 2235 623923 86773 86790 GAAATCCTGGCTAGGCCT 5-10-5 55 2236 623924 86774 86791 TGAAATCCTGGCTAGGCC 5-10-5 64 2237 623925 86775 86792 GTGAAATCCTGGCTAGGC 5-10-5 77 2238 623926 86776 86793 GGTGAAATCCTGGCTAGG 5-10-5 57 2239 623927 86777 86794 TGGTGAAATCCTGGCTAG 5-10-5 8 2240 623928 86778 86795 CTGGTGAAATCCTGGCTA 5-10-5 45 2241 623929 86780 86797 TGCTGGTGAAATCCTGGC 5-10-5 63 2242 623930 86783 86800 CACTGCTGGTGAAATCCT 5-10-5 70 2243 623931 121716 121733 ACCCTGGACCCGCCTACT 5-10-5 49 2244 623932 121721 121738 GCGCCACCCTGGACCCGC 5-10-5 84 2245 623933 121724 121741 CATGCGCCACCCTGGACC 5-10-5 52 2246 623934 121726 121743 GACATGCGCCACCCTGGA 5-10-5 80 2247 623935 121727 121744 TGACATGCGCCACCCTGG 5-10-5 74 2248 623936 121728 121745 GTGACATGCGCCACCCTG 5-10-5 90 2249 623937 121729 121746 AGTGACATGCGCCACCCT 5-10-5 84 2250 623938 121730 121747 GAGTGACATGCGCCACCC 5-10-5 92 2251 623939 121731 121748 TGAGTGACATGCGCCACC 5-10-5 86 2252 623940 121732 121749 ATGAGTGACATGCGCCAC 5-10-5 64 2253 623941 121734 121751 CGATGAGTGACATGCGCC 5-10-5 76 2254 623942 121737 121754 TTTCGATGAGTGACATGC 5-10-5 60 2255 623943 121742 121759 TCCACTTTCGATGAGTGA 5-10-5 34 2256 623944 121938 121955 CAGCACGGCGCATGGGAC 5-10-5 26 2257 623945 121939 121956 ACAGCACGGCGCATGGGA 5-10-5 62 2258 623946 121940 121957 CACAGCACGGCGCATGGG 5-10-5 57 2259 621455 121941 121958 CCACAGCACGGCGCATGG 5-10-5 44 1232 623804 121942 121959 GCCACAGCACGGCGCATG 5-10-5 85 1657 623947 121943 121960 AGCCACAGCACGGCGCAT 5-10-5 83 2260 623948 121944 121961 AAGCCACAGCACGGCGCA 5-10-5 56 2261 623949 121946 121963 TCAAGCCACAGCACGGCG 5-10-5 58 2262 623950 121949 121966 AATTCAAGCCACAGCACG 5-10-5 52 2263 623951 121954 121971 CTAATAATTCAAGCCACA 5-10-5 56 2264 623952 125424 125441 GACATTTGCTCAGCAAAC 5-10-5 74 2265 623953 125426 125443 CAGACATTTGCTCAGCAA 5-10-5 62 2266 623954 125427 125444 CCAGACATTTGCTCAGCA 5-10-5 75 2267 623955 125428 125445 CCCAGACATTTGCTCAGC 5-10-5 65 2268 623956 125429 125446 ACCCAGACATTTGCTCAG 5-10-5 30 2269 623957 125430 125447 GACCCAGACATTTGCTCA 5-10-5 30 2270 623958 125431 125448 AGACCCAGACATTTGCTC 5-10-5 41 2271 623959 125432 125449 AAGACCCAGACATTTGCT 5-10-5 69 2272 623960 125434 125451 GCAAGACCCAGACATTTG 5-10-5 73 2273 623961 125437 125454 TGTGCAAGACCCAGACAT 5-10-5 62 2274 623962 125442 125459 GTCATTGTGCAAGACCCA 5-10-5 87 2275

TABLE 49 Inhibition of tau mRNA by 5-10-5 MOE, 5-8-5 MOE, 4-8-6 MOE, and 6-8-4 gapmers targeting SEQ ID NO: 1 SEQ ID NO: 1 SEQ ID SEQ Start NO: 1 % ID ISIS NO Site Stop Site Sequence Motif inhibition NO 613412 73879 73898 GTGATCTTCCATCACTTCGA 5-10-5 87 25 620887 98891 98908 GTTTTCAAACACACCTTC 5-8-5 96 665 98928 98945 620888 98881 98898 ACACCTTCATTTACTGTC 5-8-5 96 897 98918 98935 620889 98890 98907 TTTTCAAACACACCTTCA 5-8-5 85 898 98927 98944 620890 98892 98909 GGTTTTCAAACACACCTT 5-8-5 95 899 98929 98946 620891 98893 98910 TGGTTTTCAAACACACCT 5-8-5 96 900 98930 98947 623963 73877 73894 TCTTCCATCACTTCGAAC 5-8-5 58 2276 623964 73878 73895 ATCTTCCATCACTTCGAA 5-8-5 44 2277 623965 73880 73897 TGATCTTCCATCACTTCG 5-8-5 51 2278 623966 73881 73898 GTGATCTTCCATCACTTC 5-8-5 45 2279 623967 73931 73948 ATGGTGTAGCCCCCCTGA 5-8-5 66 2280 623968 73932 73949 CATGGTGTAGCCCCCCTG 5-8-5 87 2281 623969 73934 73951 TGCATGGTGTAGCCCCCC 5-8-5 94 2282 623970 73935 73952 GTGCATGGTGTAGCCCCC 5-8-5 90 2283 623971 73957 73974 CCGTGTCACCCTCTTGGT 5-8-5 83 2284 623972 73958 73975 TCCGTGTCACCCTCTTGG 5-8-5 79 2285 623973 73960 73977 CGTCCGTGTCACCCTCTT 5-8-5 92 2286 623974 83429 83446 TCTTAGCATCAGAGGTTT 5-8-5 70 2287 623975 83430 83447 CTCTTAGCATCAGAGGTT 5-8-5 76 2288 623976 83432 83449 TGCTCTTAGCATCAGAGG 5-8-5 72 2289 623977 83433 83450 GTGCTCTTAGCATCAGAG 5-8-5 86 2290 623978 95266 95283 CTCCTCCGAGTGCGCCTG 5-8-5 24 2291 623979 95267 95284 GCTCCTCCGAGTGCGCCT 5-8-5 77 2292 623980 95269 95286 ATGCTCCTCCGAGTGCGC 5-8-5 58 2293 623981 95270 95287 AATGCTCCTCCGAGTGCG 5-8-5 79 2294 623982 95272 95289 CAAATGCTCCTCCGAGTG 5-8-5 34 2295 623983 98557 98574 TTACTGACCATGCGAGCT 5-8-5 79 2296 623984 98558 98575 TTTACTGACCATGCGAGC 5-8-5 73 2297 623985 98560 98577 CTTTTACTGACCATGCGA 5-8-5 85 2298 623986 98561 98578 GCTTTTACTGACCATGCG 5-8-5 91 2299 623987 98563 98580 TTGCTTTTACTGACCATG 5-8-5 96 2300 623988 135784 135801 ACTGCGAGGAGCAGCTGG 5-8-5 43 2301 623989 135785 135802 AACTGCGAGGAGCAGCTG 5-8-5 31 2302 623990 135787 135804 CGAACTGCGAGGAGCAGC 5-8-5 62 2303 623991 135788 135805 CCGAACTGCGAGGAGCAG 5-8-5 73 2304 623992 135790 135807 AACCGAACTGCGAGGAGC 5-8-5 73 2305 623993 135868 135885 TGCTCTTACTCCCATCAC 5-8-5 59 2306 623994 135869 135886 TTGCTCTTACTCCCATCA 5-8-5 85 2307 623995 135871 135888 ATTTGCTCTTACTCCCAT 5-8-5 77 2308 623996 135872 135889 AATTTGCTCTTACTCCCA 5-8-5 80 2309 625423 98879 98896 ACCTTCATTTACTGTCAG 5-8-5 90 2310 625424 98880 98897 CACCTTCATTTACTGTCA 5-8-5 94 2311 625425 98882 98899 CACACCTTCATTTACTGT 5-8-5 89 2312 98919 98936 5-8-5 625426 98883 98900 ACACACCTTCATTTACTG 5-8-5 87 2313 98920 98937 625427 98884 98901 AACACACCTTCATTTACT 5-8-5 86 2314 98921 98938 625428 98885 98902 AAACACACCTTCATTTAC 5-8-5 49 2315 98922 98939 625429 98886 98903 CAAACACACCTTCATTTA 5-8-5 66 2316 98923 98940 625430 98887 98904 TCAAACACACCTTCATTT 5-8-5 70 2317 98924 98941 625431 98888 98905 TTCAAACACACCTTCATT 5-8-5 74 2318 98925 98942 625432 98889 98906 TTTCAAACACACCTTCAT 5-8-5 81 2319 98926 98943 625433 98894 98911 TTGGTTTTCAAACACACC 5-8-5 92 2320 625434 75128 75145 AGCATTAAAAGAGAAAAG 5-8-5 13 2321 625435 75136 75153 TTATTTTTAGCATTAAAA 5-8-5 0 2322 625436 75138 75155 TCTTATTTTTAGCATTAA 5-8-5 52 2323 625437 75139 75156 TTCTTATTTTTAGCATTA 5-8-5 86 2324 625438 75140 75157 TTTCTTATTTTTAGCATT 5-8-5 40 2325 625439 75141 75158 TTTTCTTATTTTTAGCAT 5-8-5 0 2326 625440 75142 75159 GTTTTCTTATTTTTAGCA 5-8-5 82 2327 625441 75143 75160 TGTTTTCTTATTTTTAGC 5-8-5 68 2328 625442 75146 75163 AGGTGTTTTCTTATTTTT 5-8-5 83 2329 625443 75154 75171 GGGCTGCAAGGTGTTTTC 5-8-5 69 2330 625444 98881 98898 ACACCTTCATTTACTGTC 4-8-6 90 897 98918 98935 625445 98882 98899 CACACCTTCATTTACTGT 4-8-6 82 2312 98919 98936 625446 98883 98900 ACACACCTTCATTTACTG 4-8-6 84 2313 98920 98937 625447 98884 98901 AACACACCTTCATTTACT 4-8-6 40 2314 98921 98938 625448 98889 98906 TTTCAAACACACCTTCAT 4-8-6 83 2319 98926 98943 625449 98890 98907 TTTTCAAACACACCTTCA 4-8-6 0 898 98927 98944 625450 98891 98908 GTTTTCAAACACACCTTC 4-8-6 94 665 98928 98945 625451 98892 98909 GGTTTTCAAACACACCTT 4-8-6 94 899 98929 98946 625452 98893 98910 TGGTTTTCAAACACACCT 4-8-6 94 900 98930 98947 625453 98881 98898 ACACCTTCATTTACTGTC 6-8-4 96 897 98918 98935 625454 98882 98899 CACACCTTCATTTACTGT 6-8-4 96 2312 98919 98936 625455 98883 98900 ACACACCTTCATTTACTG 6-8-4 93 2313 98920 98937 625456 98884 98901 AACACACCTTCATTTACT 6-8-4 88 2314 98921 98938 625457 98889 98906 TTTCAAACACACCTTCAT 6-8-4 85 2319 98926 98943 625458 98890 98907 TTTTCAAACACACCTTCA 6-8-4 90 898 98927 98944 625459 98891 98908 GTTTTCAAACACACCTTC 6-8-4 97 665 98928 98945 625460 98892 98909 GGTTTTCAAACACACCTT 6-8-4 97 899 98929 98946 625461 98893 98910 TGGTTTTCAAACACACCT 6-8-4 96 900 98930 98947

Example 13 Dose-Dependent Antisense Inhibition of Human Tau in SH-SY5Y Cells by MOE Gapmers

Gapmers from studies described above exhibiting significant in vitro inhibition of tau mRNA were selected and tested at various doses in SH-SY5Y cells. The antisense oligonucleotides were tested in a series of experiments that had similar culture conditions. The results for each experiment are presented in separate tables shown below. Cells were plated at a density of 20,000 cells per well and transfected using electroporation with 0.938 μM, 0.1.875 μM, 3.750 μM, 7.500 μM, and 15.00 μM concentrations of antisense oligonucleotide, as specified in the Tables below. After a treatment period of approximately 16 hours, RNA was isolated from the cells and tau mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3104 was used to measure mRNA levels. Tau mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of tau, relative to untreated control cells. Tau mRNA levels were significantly reduced in a dose-dependent manner in antisense oligonucleotide treated cells.

TABLE 50 ISIS No 0.938 μM 1.875 μM 3.750 μM 7.500 μM 15.00 μM 613412 57 53 49 83 91 620887 68 84 95 95 97 620919 35 54 76 95 96 620930 32 49 76 86 92 620946 41 61 73 90 92 620971 61 63 75 88 96 620976 40 68 80 91 96 620978 49 49 78 93 94 620988 64 58 76 92 95 621031 51 72 90 89 92 621032 35 55 81 89 96 621041 48 73 80 88 95 621236 62 59 78 86 88 621239 46 62 63 94 94 621254 63 67 88 84 87

TABLE 51 ISIS No 0.938 μM 1.875 μM 3.750 μM 7.500 μM 15.00 μM 613412 33 54 63 89 95 620887 72 90 95 96 96 620889 20 45 72 87 95 621417 52 68 81 88 92 621434 45 64 69 84 76 621440 58 72 77 88 87 621492 40 60 80 78 83 621599 43 73 83 91 95 621615 36 67 74 81 87 621620 48 74 88 95 98 621644 4 28 33 55 65 623685 30 48 73 87 93 623872 31 61 80 90 94 623879 22 50 74 88 95 623932 39 62 80 91 95

TABLE 52 ISIS No 0.938 μM 1.875 μM 3.750 μM 7.500 μM 15.00 μM 613412 23 51 50 87 94 620887 64 82 92 95 96 620891 79 89 93 96 96 623757 38 58 73 85 95 623783 30 40 67 78 91 623804 51 70 81 87 92 623902 62 77 90 87 95 623936 46 70 84 91 94 623937 48 58 79 91 93 623938 39 63 76 90 93 623939 29 57 77 91 93 623962 44 68 78 87 91 625442 32 57 66 82 88 625459 74 87 95 96 98 625460 88 94 97 97 98

TABLE 53 ISIS No 0.938 μM 1.875 μM 3.750 μM 7.500 μM 15.00 μM 613412 6 11 20 0 35 620887 71 86 92 95 96 620888 73 89 93 95 96 620890 82 90 94 95 96 623969 41 62 84 94 n.d. 623987 46 71 89 96 97 625424 50 73 88 94 96 625433 64 83 90 95 95 625450 69 84 91 96 97 625451 85 91 95 93 95 625452 67 83 91 93 95 625453 72 85 91 94 96 625454 73 86 93 95 96 625455 44 68 86 92 94 625461 75 84 94 95 97

TABLE 54 ISIS No 0.938 μM 1.875 μM 3.750 μM 7.500 μM 15.00 μM 613412 11 30 65 82 83 620887 59 77 66 95 79 623968 43 57 72 86 93 623970 29 54 77 88 94 623973 48 65 80 93 94 623977 38 60 72 82 89 623986 45 53 80 91 96 625423 43 57 75 77 95 625425 39 49 78 88 93 625426 15 43 61 82 92 625427 16 35 64 81 93 625437 30 45 64 84 91 625444 28 45 67 84 92 625446 16 33 58 76 92 625456 27 43 57 86 91 625458 19 45 61 85 93

TABLE 55 ISIS No 0.938 μM 1.875 μM 3.750 μM 7.500 μM 15.00 μM 613412 0 10 31 71 77 620887 33 58 74 82 92 621842 7 31 45 72 87 621846 0 0 28 62 90 623903 0 0 25 71 83 623985 0 17 41 74 84 623994 19 42 67 81 83 625457 0 5 57 69 80

Example 14 Design of 5-7-6 MOE, 5-8-5 MOE, 5-9-5 MOE, and 5-10-5 MOE Gapmers with Phosphorothioate and Phosphodiester Internucleoside Linkages at a Hot Spot Region of Human Tau

Antisense oligonucleotides were designed targeting a tau nucleic acid at a region identified as a ‘hotspot’ in the studies above.

The newly designed chimeric antisense oligonucleotides in the Table below were designed as 5-7-6 MOE, 5-8-5 MOE, 5-9-5 MOE, or 5-10-5 MOE gapmers. The 5-7-6 MOE gapmers are 18 nucleosides in length, wherein the central gap segment comprises of seven 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five and six nucleosides respectively. The 5-8-5 MOE gapmers are 18 nucleosides in length, wherein the central gap segment comprises of eight 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five nucleosides each. The 5-9-5 MOE gapmers are 19 nucleosides in length, wherein the central gap segment comprises of nine 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five nucleosides each. The 5-10-5 MOE gapmers are 20 nucleosides in length, wherein the central gap segment comprises of ten 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five nucleosides each. Each nucleoside in the 5′ wing segment and each nucleoside in the 3′ wing segment has a 2′-MOE modification. The internucleoside linkages throughout each gapmer are either phosphorothioate linkages or phosphodiester linkages. The ‘Chemistry’ column describes the internucleoside linkages of each oligonucleotide. ‘s’ indicates phosphorothioate linkage and ‘o’ indicates phosphodiester linkage. All cytosine residues throughout each gapmer are 5-methylcytosines.

“Start site” indicates the 5′-most nucleoside to which the gapmer is targeted in the human gene sequence. “Stop site” indicates the 3′-most nucleoside to which the gapmer is targeted in the human gene sequence. Each gapmer listed in the Table below is targeted to either the human tau genomic sequence, designated herein as SEQ ID NO: 1 (GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to 9381000) or to the human tau mRNA sequence, designated herein as SEQ ID NO: 2 (GENBANK Accession No. NM_001123066.3). ‘n/a’ indicates that the antisense oligonucleotide does not target that particular gene sequence with 100% complementarity.

TABLE 56 MOE gapmers targeting SEQ ID NOs: 1 and 2 SEQ SEQ SEQ SEQ ID ID ID ID NO: 1 NO: 1 NO: 2 NO: 2 SEQ ISIS Start Stop Start Stop ID NO Site Site Sequence Site Site Linkage chemistry Motif NO 664511 135820 135837 GACAAAAGCAGGTTAAGT 2783 2800 sooosssssssssooss 5-8-5 2331 664714 135820 135838 TGACAAAAGCAGGTTAAGT 2783 2801 sooossssssssssooss 5-9-5 2332 664661 135820 135839 GTGACAAAAGCAGGTTAAGT 2783 2802 sooosssssssssssooss 5-10-5 2333 664767 135820 135837 GACAAAAGCAGGTTAAGT 2783 2800 sooossssssssoooss 5-7-6 2331 664512 135821 135838 TGACAAAAGCAGGTTAAG 2784 2801 sooosssssssssooss 5-8-5 2334 664715 135821 135839 GTGACAAAAGCAGGTTAAG 2784 2802 sooossssssssssooss 5-9-5 2335 664662 135821 135840 AGTGACAAAAGCAGGTTAAG 2784 2803 sooosssssssssssooss 5-10-5 2336 664768 135821 135838 TGACAAAAGCAGGTTAAG 2784 2801 sooossssssssoooss 5-7-6 2334 622109 135822 135839 GTGACAAAAGCAGGTTAA 2785 2802 sooosssssssssooss 5-8-5 2038 664716 135822 135840 AGTGACAAAAGCAGGTTAA 2785 2803 sooossssssssssooss 5-9-5 2337 664663 135822 135841 GAGTGACAAAAGCAGGTTAA 2785 2804 sooosssssssssssooss 5-10-5 2338 664769 135822 135839 GTGACAAAAGCAGGTTAA 2785 2802 sooossssssssoooss 5-7-6 2038 664513 135823 135840 AGTGACAAAAGCAGGTTA 2786 2803 sooosssssssssooss 5-8-5 2339 664717 135823 135841 GAGTGACAAAAGCAGGTTA 2786 2804 sooossssssssssooss 5-9-5 2340 664664 135823 135842 CGAGTGACAAAAGCAGGTTA 2786 2805 sooosssssssssssooss 5-10-5 2341 664770 135823 135840 AGTGACAAAAGCAGGTTA 2786 2803 sooossssssssoooss 5-7-6 2339 664514 135824 135841 GAGTGACAAAAGCAGGTT 2787 2804 sooosssssssssooss 5-8-5 2342 664718 135824 135842 CGAGTGACAAAAGCAGGTT 2787 2805 sooossssssssssooss 5-9-5 2343 664665 135824 135843 CCGAGTGACAAAAGCAGGTT 2787 2806 sooosssssssssssooss 5-10-5 2344 664771 135824 135841 GAGTGACAAAAGCAGGTT 2787 2804 sooossssssssoooss 5-7-6 2342 622110 135825 135842 CGAGTGACAAAAGCAGGT 2788 2805 sooosssssssssooss 5-8-5 2039 664719 135825 135843 CCGAGTGACAAAAGCAGGT 2788 2806 sooossssssssssooss 5-9-5 2345 664666 135825 135844 GCCGAGTGACAAAAGCAGGT 2788 2807 sooosssssssssssooss 5-10-5 2346 664772 135825 135842 CGAGTGACAAAAGCAGGT 2788 2805 sooossssssssoooss 5-7-6 2039 664515 135826 135843 CCGAGTGACAAAAGCAGG 2789 2806 sooosssssssssooss 5-8-5 2347 664720 135826 135844 GCCGAGTGACAAAAGCAGG 2789 2807 sooossssssssssooss 5-9-5 2348 664667 135826 135845 AGCCGAGTGACAAAAGCAGG 2789 2808 sooosssssssssssooss 5-10-5 2349 664773 135826 135843 CCGAGTGACAAAAGCAGG 2789 2806 sooossssssssoooss 5-7-6 2347 664516 135827 135844 GCCGAGTGACAAAAGCAG 2790 2807 sooosssssssssooss 5-8-5 2350 664721 135827 135845 AGCCGAGTGACAAAAGCAG 2790 2808 sooossssssssssooss 5-9-5 2351 664668 135827 135846 AAGCCGAGTGACAAAAGCAG 2790 2809 sooosssssssssssooss 5-10-5 2352 664774 135827 135844 GCCGAGTGACAAAAGCAG 2790 2807 sooossssssssoooss 5-7-6 2350 622111 135828 135845 AGCCGAGTGACAAAAGCA 2791 2808 sooosssssssssooss 5-8-5 2040 664722 135828 135846 AAGCCGAGTGACAAAAGCA 2791 2809 sooossssssssssooss 5-9-5 2353 664669 135828 135847 AAAGCCGAGTGACAAAAGCA 2791 2810 sooosssssssssssooss 5-10-5 2354 664775 135828 135845 AGCCGAGTGACAAAAGCA 2791 2808 sooossssssssoooss 5-7-6 2040 664517 135829 135846 AAGCCGAGTGACAAAAGC 2792 2809 sooosssssssssooss 5-8-5 2355 664723 135829 135847 AAAGCCGAGTGACAAAAGC 2792 2810 sooossssssssssooss 5-9-5 2356 664670 135829 135848 CAAAGCCGAGTGACAAAAGC 2792 2811 sooosssssssssssooss 5-10-5 2357 664776 135829 135846 AAGCCGAGTGACAAAAGC 2792 2809 sooossssssssoooss 5-7-6 2355 664518 135830 135847 AAAGCCGAGTGACAAAAG 2793 2810 sooosssssssssooss 5-8-5 2358 664724 135830 135848 CAAAGCCGAGTGACAAAAG 2793 2811 sooossssssssssooss 5-9-5 2359 664671 135830 135849 CCAAAGCCGAGTGACAAAAG 2793 2812 sooosssssssssssooss 5-10-5 2360 664777 135830 135847 AAAGCCGAGTGACAAAAG 2793 2810 sooossssssssoooss 5-7-6 2358 622112 135831 135848 CAAAGCCGAGTGACAAAA 2794 2811 sooosssssssssooss 5-8-5 2041 664725 135831 135849 CCAAAGCCGAGTGACAAAA 2794 2812 sooossssssssssooss 5-9-5 2361 664672 135831 135850 GCCAAAGCCGAGTGACAAAA 2794 2813 sooosssssssssssooss 5-10-5 2362 664778 135831 135848 CAAAGCCGAGTGACAAAA 2794 2811 sooossssssssoooss 5-7-6 2041 664519 135832 135849 CCAAAGCCGAGTGACAAA 2795 2812 sooosssssssssooss 5-8-5 2363 664726 135832 135850 GCCAAAGCCGAGTGACAAA 2795 2813 sooossssssssssooss 5-9-5 2364 664673 135832 135851 AGCCAAAGCCGAGTGACAAA 2795 2814 sooosssssssssssooss 5-10-5 2365 664779 135832 135849 CCAAAGCCGAGTGACAAA 2795 2812 sooossssssssoooss 5-7-6 2363 664520 135833 135850 GCCAAAGCCGAGTGACAA 2796 2813 sooosssssssssooss 5-8-5 2366 664727 135833 135851 AGCCAAAGCCGAGTGACAA 2796 2814 sooossssssssssooss 5-9-5 2367 664674 135833 135852 GAGCCAAAGCCGAGTGACAA 2796 2815 sooosssssssssssooss 5-10-5 2368 664780 135833 135850 GCCAAAGCCGAGTGACAA 2796 2813 sooossssssssoooss 5-7-6 2366 622113 135834 135851 AGCCAAAGCCGAGTGACA 2797 2814 sooosssssssssooss 5-8-5 2042 664728 135834 135852 GAGCCAAAGCCGAGTGACA 2797 2815 sooossssssssssooss 5-9-5 2369 664675 135834 135853 CGAGCCAAAGCCGAGTGACA 2797 2816 sooosssssssssssooss 5-10-5 2419 664781 135834 135851 AGCCAAAGCCGAGTGACA 2797 2814 sooossssssssoooss 5-7-6 2042 664521 135835 135852 GAGCCAAAGCCGAGTGAC 2798 2815 sooosssssssssooss 5-8-5 2420 664729 135835 135853 CGAGCCAAAGCCGAGTGAC 2798 2816 sooossssssssssooss 5-9-5 2421 664676 135835 135854 CCGAGCCAAAGCCGAGTGAC 2798 2817 sooosssssssssssooss 5-10-5 2422 664782 135835 135852 GAGCCAAAGCCGAGTGAC 2798 2815 sooossssssssoooss 5-7-6 2420 664522 135836 135853 CGAGCCAAAGCCGAGTGA 2799 2816 sooosssssssssooss 5-8-5 2423 664730 135836 135854 CCGAGCCAAAGCCGAGTGA 2799 2817 sooossssssssssooss 5-9-5 2424 664677 135836 135855 CCCGAGCCAAAGCCGAGTGA 2799 2818 sooosssssssssssooss 5-10-5 2425 664783 135836 135853 CGAGCCAAAGCCGAGTGA 2799 2816 sooossssssssoooss 5-7-6 2423 622114 135837 135854 CCGAGCCAAAGCCGAGTG 2800 2817 sooosssssssssooss 5-8-5 2043 664731 135837 135855 CCCGAGCCAAAGCCGAGTG 2800 2818 sooossssssssssooss 5-9-5 2426 664678 135837 135856 TCCCGAGCCAAAGCCGAGTG 2800 2819 sooosssssssssssooss 5-10-5 2427 664784 135837 135854 CCGAGCCAAAGCCGAGTG 2800 2817 sooossssssssoooss 5-7-6 2043 664523 135838 135855 CCCGAGCCAAAGCCGAGT 2801 2818 sooosssssssssooss 5-8-5 2428 664732 135838 135856 TCCCGAGCCAAAGCCGAGT 2801 2819 sooossssssssssooss 5-9-5 2429 664679 135838 135857 GTCCCGAGCCAAAGCCGAGT 2801 2820 sooosssssssssssooss 5-10-5 2430 664785 135838 135855 CCCGAGCCAAAGCCGAGT 2801 2818 sooossssssssoooss 5-7-6 2428 664524 135839 135856 TCCCGAGCCAAAGCCGAG 2802 2819 sooosssssssssooss 5-8-5 2431 664733 135839 135857 GTCCCGAGCCAAAGCCGAG 2802 2820 sooossssssssssooss 5-9-5 2432 664680 135839 135858 AGTCCCGAGCCAAAGCCGAG 2802 2821 sooosssssssssssooss 5-10-5 2433 664786 135839 135856 TCCCGAGCCAAAGCCGAG 2802 2819 sooossssssssoooss 5-7-6 2431 622115 135840 135857 GTCCCGAGCCAAAGCCGA 2803 2820 sooosssssssssooss 5-8-5 2044 664734 135840 135858 AGTCCCGAGCCAAAGCCGA 2803 2821 sooossssssssssooss 5-9-5 2434 664681 135840 135859 AAGTCCCGAGCCAAAGCCGA 2803 2822 sooosssssssssssooss 5-10-5 2435 664787 135840 135857 GTCCCGAGCCAAAGCCGA 2803 2820 sooossssssssoooss 5-7-6 2044 664525 135841 135858 AGTCCCGAGCCAAAGCCG 2804 2821 sooosssssssssooss 5-8-5 2436 664735 135841 135859 AAGTCCCGAGCCAAAGCCG 2804 2822 sooossssssssssooss 5-9-5 2437 664682 135841 135860 GAAGTCCCGAGCCAAAGCCG 2804 2823 sooosssssssssssooss 5-10-5 2438 664788 135841 135858 AGTCCCGAGCCAAAGCCG 2804 2821 sooossssssssoooss 5-7-6 2436 664526 135842 135859 AAGTCCCGAGCCAAAGCC 2805 2822 sooosssssssssooss 5-8-5 2439 664736 135842 135860 GAAGTCCCGAGCCAAAGCC 2805 2823 sooossssssssssooss 5-9-5 2440 664683 135842 135861 TGAAGTCCCGAGCCAAAGCC 2805 2824 sooosssssssssssooss 5-10-5 2441 664789 135842 135859 AAGTCCCGAGCCAAAGCC 2805 2822 sooossssssssoooss 5-7-6 2439 622116 135843 135860 GAAGTCCCGAGCCAAAGC 2806 2823 sooosssssssssooss 5-8-5 2045 664737 135843 135861 TGAAGTCCCGAGCCAAAGC 2806 2824 sooossssssssssooss 5-9-5 2442 664684 135843 135862 TTGAAGTCCCGAGCCAAAGC 2806 2825 sooosssssssssssooss 5-10-5 2443 664790 135843 135860 GAAGTCCCGAGCCAAAGC 2806 2823 sooossssssssoooss 5-7-6 2045 664527 135844 135861 TGAAGTCCCGAGCCAAAG 2807 2824 sooosssssssssooss 5-8-5 2478 664738 135844 135862 TTGAAGTCCCGAGCCAAAG 2807 2825 sooossssssssssooss 5-9-5 2532 664685 135844 135863 TTTGAAGTCCCGAGCCAAAG 2807 2826 sooosssssssssssooss 5-10-5 2533 664791 135844 135861 TGAAGTCCCGAGCCAAAG 2807 2824 sooossssssssoooss 5-7-6 2478 664528 135845 135862 TTGAAGTCCCGAGCCAAA 2808 2825 sooosssssssssooss 5-8-5 2479 664739 135845 135863 TTTGAAGTCCCGAGCCAAA 2808 2826 sooossssssssssooss 5-9-5 2534 664686 135845 135864 TTTTGAAGTCCCGAGCCAAA 2808 2827 sooosssssssssssooss 5-10-5 2535 664792 135845 135862 TTGAAGTCCCGAGCCAAA 2808 2825 sooossssssssoooss 5-7-6 2479 622117 135846 135863 TTTGAAGTCCCGAGCCAA 2809 2826 sooosssssssssooss 5-8-5 2046 664740 135846 135864 TTTTGAAGTCCCGAGCCAA 2809 2827 sooossssssssssooss 5-9-5 2536 664687 135846 135865 ATTTTGAAGTCCCGAGCCAA 2809 2828 sooosssssssssssooss 5-10-5 2537 664793 135846 135863 TTTGAAGTCCCGAGCCAA 2809 2826 sooossssssssoooss 5-7-6 2538 664529 135847 135864 TTTTGAAGTCCCGAGCCA 2810 2827 sooosssssssssooss 5-8-5 2480 664741 135847 135865 ATTTTGAAGTCCCGAGCCA 2810 2828 sooossssssssssooss 5-9-5 2539 664688 135847 135866 GATTTTGAAGTCCCGAGCCA 2810 2829 sooosssssssssssooss 5-10-5 2540 664794 135847 135864 TTTTGAAGTCCCGAGCCA 2810 2827 sooossssssssoooss 5-7-6 2480 664530 135848 135865 ATTTTGAAGTCCCGAGCC 2811 2828 sooosssssssssooss 5-8-5 2481 664742 135848 135866 GATTTTGAAGTCCCGAGCC 2811 2829 sooossssssssssooss 5-9-5 2541 664689 135848 135867 TGATTTTGAAGTCCCGAGCC 1644 1663 sooosssssssssssooss 5-10-5 56 664795 135848 135865 ATTTTGAAGTCCCGAGCC 2811 2828 sooossssssssoooss 5-7-6 2481 622118 135849 135866 GATTTTGAAGTCCCGAGC 2812 2829 sooosssssssssooss 5-8-5 2047 664743 135849 135867 TGATTTTGAAGTCCCGAGC 2812 2830 sooossssssssssooss 5-9-5 2542 664690 135849 135868 CTGATTTTGAAGTCCCGAGC 2812 2831 sooosssssssssssooss 5-10-5 464 664796 135849 135866 GATTTTGAAGTCCCGAGC 2812 2829 sooossssssssoooss 5-7-6 2047 664531 135850 135867 TGATTTTGAAGTCCCGAG 2813 2830 sooosssssssssooss 5-8-5 2482 664744 135850 135868 CTGATTTTGAAGTCCCGAG 2813 2831 sooossssssssssooss 5-9-5 2543 664691 135850 135869 ACTGATTTTGAAGTCCCGAG 2813 2832 sooosssssssssssooss 5-10-5 2544 664797 135850 135867 TGATTTTGAAGTCCCGAG 2813 2830 sooossssssssoooss 5-7-6 2482 664532 135851 135868 CTGATTTTGAAGTCCCGA 2814 2831 sooosssssssssooss 5-8-5 2483 664745 135851 135869 ACTGATTTTGAAGTCCCGA 2814 2832 sooossssssssssooss 5-9-5 2545 664692 135851 135870 CACTGATTTTGAAGTCCCGA 2814 2833 sooosssssssssssooss 5-10-5 2546 664798 135851 135868 CTGATTTTGAAGTCCCGA 2814 2831 sooossssssssoooss 5-7-6 2483 622119 135852 135869 ACTGATTTTGAAGTCCCG 2815 2832 sooosssssssssooss 5-8-5 2048 664746 135852 135870 CACTGATTTTGAAGTCCCG 2815 2833 sooossssssssssooss 5-9-5 2547 664693 135852 135871 TCACTGATTTTGAAGTCCCG 2815 2834 sooosssssssssssooss 5-10-5 2548 664799 135852 135869 ACTGATTTTGAAGTCCCG 2815 2832 sooossssssssoooss 5-7-6 2549 664533 135853 135870 CACTGATTTTGAAGTCCC 2816 2833 sooosssssssssooss 5-8-5 2370 664747 135853 135871 TCACTGATTTTGAAGTCCC 2816 2834 sooossssssssssooss 5-9-5 2371 664694 135853 135872 ATCACTGATTTTGAAGTCCC 1649 1668 sooosssssssssssooss 5-10-5 57 664800 135853 135870 CACTGATTTTGAAGTCCC 2816 2833 sooossssssssoooss 5-7-6 2370 664534 135854 135871 TCACTGATTTTGAAGTCC 2817 2834 sooosssssssssooss 5-8-5 2372 664748 135854 135872 ATCACTGATTTTGAAGTCC 2817 2835 sooossssssssssooss 5-9-5 2373 664695 135854 135873 CATCACTGATTTTGAAGTCC 2817 2836 sooosssssssssssooss 5-10-5 2374 664801 135854 135871 TCACTGATTTTGAAGTCC 2817 2834 sooossssssssoooss 5-7-6 2372 622120 135855 135872 ATCACTGATTTTGAAGTC 2818 2835 sooosssssssssooss 5-8-5 1668 664749 135855 135873 CATCACTGATTTTGAAGTC 2818 2836 sooossssssssssooss 5-9-5 2375 664696 135855 135874 CCATCACTGATTTTGAAGTC 2818 2837 sooosssssssssssooss 5-10-5 2376 664802 135855 135872 ATCACTGATTTTGAAGTC 2818 2835 sooossssssssoooss 5-7-6 1668 664535 135856 135873 CATCACTGATTTTGAAGT 2819 2836 sooosssssssssooss 5-8-5 2377 664750 135856 135874 CCATCACTGATTTTGAAGT 2819 2837 sooossssssssssooss 5-9-5 2378 664697 135856 135875 CCCATCACTGATTTTGAAGT 2819 2838 sooosssssssssssooss 5-10-5 2379 664803 135856 135873 CATCACTGATTTTGAAGT 2819 2836 sooossssssssoooss 5-7-6 2377 664536 135857 135874 CCATCACTGATTTTGAAG 2820 2837 sooosssssssssooss 5-8-5 2380 664751 135857 135875 CCCATCACTGATTTTGAAG 2820 2838 sooossssssssssooss 5-9-5 2381 664698 135857 135876 TCCCATCACTGATTTTGAAG 2820 2839 sooosssssssssssooss 5-10-5 2382 664804 135857 135874 CCATCACTGATTTTGAAG 2820 2837 sooossssssssoooss 5-7-6 2380 622121 135858 135875 CCCATCACTGATTTTGAA 2821 2838 sooosssssssssooss 5-8-5 1669 664752 135858 135876 TCCCATCACTGATTTTGAA 2821 2839 sooossssssssssooss 5-9-5 2383 664699 135858 135877 CTCCCATCACTGATTTTGAA 2821 2840 sooosssssssssssooss 5-10-5 2384 664805 135858 135875 CCCATCACTGATTTTGAA 2821 2838 sooossssssssoooss 5-7-6 1669 664537 135859 135876 TCCCATCACTGATTTTGA 2822 2839 sooosssssssssooss 5-8-5 2385 664753 135859 135877 CTCCCATCACTGATTTTGA 2822 2840 sooossssssssssooss 5-9-5 2386 664700 135859 135878 ACTCCCATCACTGATTTTGA 2822 2841 sooosssssssssssooss 5-10-5 2387 664806 135859 135876 TCCCATCACTGATTTTGA 2822 2839 sooossssssssoooss 5-7-6 2385 664538 135860 135877 CTCCCATCACTGATTTTG 2823 2840 sooosssssssssooss 5-8-5 2388 664754 135860 135878 ACTCCCATCACTGATTTTG 2823 2841 sooossssssssssooss 5-9-5 2389 664701 135860 135879 TACTCCCATCACTGATTTTG 2823 2842 sooosssssssssssooss 5-10-5 2390 664807 135860 135877 CTCCCATCACTGATTTTG 2823 2840 sooossssssssoooss 5-7-6 2388 622122 135861 135878 ACTCCCATCACTGATTTT 2824 2841 sooosssssssssooss 5-8-5 1670 664755 135861 135879 TACTCCCATCACTGATTTT 2824 2842 sooossssssssssooss 5-9-5 2391 664702 135861 135880 TTACTCCCATCACTGATTTT 2824 2843 sooosssssssssssooss 5-10-5 2392 664808 135861 135878 ACTCCCATCACTGATTTT 2824 2841 sooossssssssoooss 5-7-6 1670 664539 135862 135879 TACTCCCATCACTGATTT 2825 2842 sooosssssssssooss 5-8-5 2393 664756 135862 135880 TTACTCCCATCACTGATTT 2825 2843 sooossssssssssooss 5-9-5 2394 664703 135862 135881 CTTACTCCCATCACTGATTT 2825 2844 sooosssssssssssooss 5-10-5 2395 664809 135862 135879 TACTCCCATCACTGATTT 2825 2842 sooossssssssoooss 5-7-6 2393 664540 135863 135880 TTACTCCCATCACTGATT 2826 2843 sooosssssssssooss 5-8-5 2396 664757 135863 135881 CTTACTCCCATCACTGATT 2826 2844 sooossssssssssooss 5-9-5 2397 664704 135863 135882 TCTTACTCCCATCACTGATT 2826 2845 sooosssssssssssooss 5-10-5 2398 664810 135863 135880 TTACTCCCATCACTGATT 2826 2843 sooossssssssoooss 5-7-6 2396 622123 135864 135881 CTTACTCCCATCACTGAT 2827 2844 sooosssssssssooss 5-8-5 1671 664758 135864 135882 TCTTACTCCCATCACTGAT 2827 2845 sooossssssssssooss 5-9-5 2399 664705 135864 135883 CTCTTACTCCCATCACTGAT 2827 2846 sooosssssssssssooss 5-10-5 2400 664811 135864 135881 CTTACTCCCATCACTGAT 2827 2844 sooossssssssoooss 5-7-6 1671 664541 135865 135882 TCTTACTCCCATCACTGA 2828 2845 sooosssssssssooss 5-8-5 2401 664759 135865 135883 CTCTTACTCCCATCACTGA 2828 2846 sooossssssssssooss 5-9-5 2402 664706 135865 135884 GCTCTTACTCCCATCACTGA 2828 2847 sooosssssssssssooss 5-10-5 2403 664812 135865 135882 TCTTACTCCCATCACTGA 2828 2845 sooossssssssoooss 5-7-6 2401 664542 135866 135883 CTCTTACTCCCATCACTG 2829 2846 sooosssssssssooss 5-8-5 2404 664760 135866 135884 GCTCTTACTCCCATCACTG 2829 2847 sooossssssssssooss 5-9-5 2405 664707 135866 135885 TGCTCTTACTCCCATCACTG 2829 2848 sooosssssssssssooss 5-10-5 2406 664813 135866 135883 CTCTTACTCCCATCACTG 2829 2846 sooossssssssoooss 5-7-6 2404 622124 135867 135884 GCTCTTACTCCCATCACT 2830 2847 sooosssssssssooss 5-8-5 1672 664761 135867 135885 TGCTCTTACTCCCATCACT 2830 2848 sooossssssssssooss 5-9-5 2407 664708 135867 135886 TTGCTCTTACTCCCATCACT 2830 2849 sooosssssssssssooss 5-10-5 2408 664814 135867 135884 GCTCTTACTCCCATCACT 2830 2847 sooossssssssoooss 5-7-6 1672 623993 135868 135885 TGCTCTTACTCCCATCAC 2831 2848 sooosssssssssooss 5-8-5 2306 664762 135868 135886 TTGCTCTTACTCCCATCAC 2831 2849 sooossssssssssooss 5-9-5 2409 664709 135868 135887 TTTGCTCTTACTCCCATCAC 2831 2850 sooosssssssssssooss 5-10-5 2410 664815 135868 135885 TGCTCTTACTCCCATCAC 2831 2848 sooossssssssoooss 5-7-6 2306 623994 135869 135886 TTGCTCTTACTCCCATCA 2832 2849 sooosssssssssooss 5-8-5 2307 664763 135869 135887 TTTGCTCTTACTCCCATCA 2832 2850 sooossssssssssooss 5-9-5 2411 664710 135869 135888 ATTTGCTCTTACTCCCATCA 2832 2851 sooosssssssssssooss 5-10-5 2412 664816 135869 135886 TTGCTCTTACTCCCATCA 2832 2849 sooossssssssoooss 5-7-6 2307 622125 135870 135887 TTTGCTCTTACTCCCATC 2833 2850 sooosssssssssooss 5-8-5 1673 664764 135870 135888 ATTTGCTCTTACTCCCATC 2833 2851 sooossssssssssooss 5-9-5 2413 664711 135870 135889 AATTTGCTCTTACTCCCATC 2833 2852 sooosssssssssssooss 5-10-5 2414 664817 135870 135887 TTTGCTCTTACTCCCATC 2833 2850 sooossssssssoooss 5-7-6 1673 623995 135871 135888 ATTTGCTCTTACTCCCAT 2834 2851 sooosssssssssooss 5-8-5 2308 664765 135871 135889 AATTTGCTCTTACTCCCAT 2834 2852 sooossssssssssooss 5-9-5 2415 664712 135871 135890 AAATTTGCTCTTACTCCCAT 2834 2853 sooosssssssssssooss 5-10-5 2416 664818 135871 135888 ATTTGCTCTTACTCCCAT 2834 2851 sooossssssssoooss 5-7-6 2308 623996 135872 135889 AATTTGCTCTTACTCCCA 2835 2852 sooosssssssssooss 5-8-5 2309 664766 135872 135890 AAATTTGCTCTTACTCCCA 2835 2853 sooossssssssssooss 5-9-5 2417 664713 135872 135891 GAAATTTGCTCTTACTCCCA 2835 2854 sooosssssssssssooss 5-10-5 2418 664819 135872 135889 AATTTGCTCTTACTCCCA 2835 2852 sooossssssssoooss 5-7-6 2309

Example 15 Intracerebroventricular Administration of Antisense Oligonucleotides Against Human Tau mRNA in Htau Mice

Selected compounds were tested for efficacy by ICV administration in human tau transgenic mice (Duff et al., Neurobiology of Disease 7:87-98, 2000).

Treatment and Surgery

Groups of 4 mice each were administered ISIS 613255, ISIS 613329, ISIS 613344, ISIS 613361, ISIS 613369, ISIS 613370, ISIS 613397, ISIS 613045, ISIS 613099, ISIS 613118, ISIS 613136 with a 200 μg dose delivered by ICV bolus injection. A control group of 2 mice was similarly treated with ISIS 424880 and a control group of 4 mice was similarly treated with PBS. All procedures were performed under isoflourane anesthesia and in accordance with IACUC regulations. For mouse ICV bolus injections, the antisense oligonucleotide was injected into the right lateral ventricle of human tau transgenic mice. Ten microliters of solution containing 300 μg of oligonucleotide in PBS was injected over approximately 10 seconds. Tissue was collected 14 days after oligonucleotide administration.

RNA Analysis

On day 14 after the oligonucleotide administration, RNA was extracted from the hippocampus, spinal cord and cortex for real-time PCR analysis of tau mRNA levels. Human tau mRNA levels were measured using the human primer probe set RTS3104. Results were calculated as percent inhibition of human tau mRNA expression compared to the control. All the antisense oligonucleotides effect significant inhibition of human tau mRNA levels.

TABLE 57 Percent reduction of human tau mRNA levels in hTau mice ISIS Spinal No Cortex Hippocampus Cord 613255 30 46 36 613329 20 69 67 613344 41 42 34 613361 69 57 72 613369 17 48 46 613370 42 61 63 613397 33 41 57 613045 12 38 47 613099 42 55 54 613118 64 73 58 613136 26 39 27 424880 50 53 55

Example 16 Antisense Inhibition of Human Tau in SH-SY5Y Cells 5-7-6 MOE, 5-8-5 MOE, 5-9-5 MOE, and 5-10-5 MOE Gapmers

The antisense oligonucleotides described in the Examples above, as well as newly designed antisense oligonucleotides targeting a human tau nucleic acid, were tested in a series of experiments that had similar culture conditions. The results for each experiment are presented in separate tables shown below.

Cultured SH-SY5Y cells were transfected using electroporation with 8,000 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and tau mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3104 was used to measure mRNA levels. Tau mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of tau, relative to untreated control cells.

The newly designed chimeric antisense oligonucleotides in the Tables below were designed as 5-7-6 MOE, 5-8-5 MOE, 5-9-5 MOE, or 5-10-5 MOE gapmers. The 5-7-6 MOE gapmers are 18 nucleosides in length, wherein the central gap segment comprises of seven 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five and six nucleosides respectively. The 5-8-5 MOE gapmers are 18 nucleosides in length, wherein the central gap segment comprises of eight 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five nucleosides each. The 5-9-5 MOE gapmers are 19 nucleosides in length, wherein the central gap segment comprises of nine 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five nucleosides each. The 5-10-5 MOE gapmers are 20 nucleosides in length, wherein the central gap segment comprises of ten 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five nucleosides each. Each nucleoside in the 5′ wing segment and each nucleoside in the 3′ wing segment has a 2′-MOE modification. The internucleoside linkages throughout each gapmer are either phosphorothioate linkages or phosphodiester linkages. The ‘Linkage Chemistry’ column describes the internucleoside linkages of each oligonucleotide. ‘s’ indicates phosphorothioate linkage and ‘o’ indicates phosphodiester linkage. All cytosine residues throughout each gapmer are 5-methylcytosines.

“Start site” indicates the 5′-most nucleoside to which the gapmer is targeted in the human gene sequence. “Stop site” indicates the 3′-most nucleoside to which the gapmer is targeted in the human gene sequence. Each gapmer listed in the Table below is targeted to either the human tau genomic sequence, designated herein as SEQ ID NO: 1 (GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to 9381000) or to the human tau mRNA sequence, designated herein as SEQ ID NO: 2 (GENBANK Accession No. NM_001123066.3). ‘n/a’ indicates that the antisense oligonucleotide does not target that particular gene sequence with 100% complementarity.

TABLE 58 SEQ SEQ SEQ SEQ ID ID ID ID NO: 1 NO: 1 NO: 2 NO: 2 SEQ ISIS start stop start stop Linkage % ID NO site site Sequence site site Chemistry Motif inhibition NO 623965 73880 73897 TGATCTTCCATCACTTCG n/a n/a sooosssssssssooss 5-8-5 52 2278 620887 98891 98908 GTTTTCAAACACACCTTC n/a n/a sooosssssssssooss 5-8-5 91 665 98928 98945 664511 135820 135837 GACAAAAGCAGGTTAAGT 2783 2800 sooosssssssssooss 5-8-5 65 2331 664767 135820 135837 GACAAAAGCAGGTTAAGT 2783 2800 sooossssssssoooss 5-7-6 21 2331 664512 135821 135838 TGACAAAAGCAGGTTAAG 2784 2801 sooosssssssssooss 5-8-5 60 2334 664768 135821 135838 TGACAAAAGCAGGTTAAG 2784 2801 sooossssssssoooss 5-7-6 30 2334 622109 135822 135839 GTGACAAAAGCAGGTTAA 2785 2802 sooosssssssssooss 5-8-5 72 2038 664769 135822 135839 GTGACAAAAGCAGGTTAA 2785 2802 sooossssssssoooss 5-7-6 49 2038 664513 135823 135840 AGTGACAAAAGCAGGTTA 2786 2803 sooosssssssssooss 5-8-5 56 2339 664770 135823 135840 AGTGACAAAAGCAGGTTA 2786 2803 sooossssssssoooss 5-7-6 45 2339 664514 135824 135841 GAGTGACAAAAGCAGGTT 2787 2804 sooosssssssssooss 5-8-5 84 2342 664771 135824 135841 GAGTGACAAAAGCAGGTT 2787 2804 sooossssssssoooss 5-7-6 63 2342 622110 135825 135842 CGAGTGACAAAAGCAGGT 2788 2805 sooosssssssssooss 5-8-5 77 2039 664772 135825 135842 CGAGTGACAAAAGCAGGT 2788 2805 sooossssssssoooss 5-7-6 37 2039 664515 135826 135843 CCGAGTGACAAAAGCAGG 2789 2806 sooosssssssssooss 5-8-5 70 2347 664773 135826 135843 CCGAGTGACAAAAGCAGG 2789 2806 sooossssssssoooss 5-7-6 62 2347 664516 135827 135844 GCCGAGTGACAAAAGCAG 2790 2807 sooosssssssssooss 5-8-5 79 2350 664774 135827 135844 GCCGAGTGACAAAAGCAG 2790 2807 sooossssssssoooss 5-7-6 70 2350 622111 135828 135845 AGCCGAGTGACAAAAGCA 2791 2808 sooosssssssssooss 5-8-5 75 2040 664775 135828 135845 AGCCGAGTGACAAAAGCA 2791 2808 sooossssssssoooss 5-7-6 78 2040 664517 135829 135846 AAGCCGAGTGACAAAAGC 2792 2809 sooosssssssssooss 5-8-5 67 2355 664776 135829 135846 AAGCCGAGTGACAAAAGC 2792 2809 sooossssssssoooss 5-7-6 50 2355 664518 135830 135847 AAAGCCGAGTGACAAAAG 2793 2810 sooosssssssssooss 5-8-5 47 2358 664777 135830 135847 AAAGCCGAGTGACAAAAG 2793 2810 sooossssssssoooss 5-7-6 42 2358 622112 135831 135848 CAAAGCCGAGTGACAAAA 2794 2811 sooosssssssssooss 5-8-5 46 2041 664778 135831 135848 CAAAGCCGAGTGACAAAA 2794 2811 sooossssssssoooss 5-7-6 34 2041 664519 135832 135849 CCAAAGCCGAGTGACAAA 2795 2812 sooosssssssssooss 5-8-5 57 2363 664779 135832 135849 CCAAAGCCGAGTGACAAA 2795 2812 sooossssssssoooss 5-7-6 34 2363 664520 135833 135850 GCCAAAGCCGAGTGACAA 2796 2813 sooosssssssssooss 5-8-5 63 2366 664780 135833 135850 GCCAAAGCCGAGTGACAA 2796 2813 sooossssssssoooss 5-7-6 66 2366 622113 135834 135851 AGCCAAAGCCGAGTGACA 2797 2814 sooosssssssssooss 5-8-5 70 2042 664781 135834 135851 AGCCAAAGCCGAGTGACA 2797 2814 sooossssssssoooss 5-7-6 74 2042 664521 135835 135852 GAGCCAAAGCCGAGTGAC 2798 2815 sooosssssssssooss 5-8-5 71 2420 664782 135835 135852 GAGCCAAAGCCGAGTGAC 2798 2815 sooossssssssoooss 5-7-6 31 2420 664522 135836 135853 CGAGCCAAAGCCGAGTGA 2799 2816 sooosssssssssooss 5-8-5 50 2423 664783 135836 135853 CGAGCCAAAGCCGAGTGA 2799 2816 sooossssssssoooss 5-7-6 30 2423 622114 135837 135854 CCGAGCCAAAGCCGAGTG 2800 2817 sooosssssssssooss 5-8-5 72 2043 664784 135837 135854 CCGAGCCAAAGCCGAGTG 2800 2817 sooossssssssoooss 5-7-6 76 2043 664523 135838 135855 CCCGAGCCAAAGCCGAGT 2801 2818 sooosssssssssooss 5-8-5 70 2428 664785 135838 135855 CCCGAGCCAAAGCCGAGT 2801 2818 sooossssssssoooss 5-7-6 56 2428 664524 135839 135856 TCCCGAGCCAAAGCCGAG 2802 2819 sooosssssssssooss 5-8-5 55 2431 664786 135839 135856 TCCCGAGCCAAAGCCGAG 2802 2819 sooossssssssoooss 5-7-6 36 2431 622115 135840 135857 GTCCCGAGCCAAAGCCGA 2803 2820 sooosssssssssooss 5-8-5 59 2044 664787 135840 135857 GTCCCGAGCCAAAGCCGA 2803 2820 sooossssssssoooss 5-7-6 54 2044 664525 135841 135858 AGTCCCGAGCCAAAGCCG 2804 2821 sooosssssssssooss 5-8-5 74 2436 664788 135841 135858 AGTCCCGAGCCAAAGCCG 2804 2821 sooossssssssoooss 5-7-6 77 2436 664789 135842 135859 AAGTCCCGAGCCAAAGCC 2805 2822 sooossssssssoooss 5-7-6 42 2439 664790 135843 135860 GAAGTCCCGAGCCAAAGC 2806 2823 sooossssssssoooss 5-7-6 38 2045 664791 135844 135861 TGAAGTCCCGAGCCAAAG 2807 2824 sooossssssssoooss 5-7-6 44 2478 664792 135845 135862 TTGAAGTCCCGAGCCAAA 2808 2825 sooossssssssoooss 5-7-6 44 2479 664793 135846 135863 TTTGAAGTCCCGAGCCAA 2809 2826 sooossssssssoooss 5-7-6 29 2046 664794 135847 135864 TTTTGAAGTCCCGAGCCA 2810 2827 sooossssssssoooss 5-7-6 32 2480 664795 135848 135865 ATTTTGAAGTCCCGAGCC 2811 2828 sooossssssssoooss 5-7-6 15 2481 664796 135849 135866 GATTTTGAAGTCCCGAGC 2812 2829 sooossssssssoooss 5-7-6 76 2047 664797 135850 135867 TGATTTTGAAGTCCCGAG 2813 2830 sooossssssssoooss 5-7-6 70 2482 664798 135851 135868 CTGATTTTGAAGTCCCGA 2814 2831 sooossssssssoooss 5-7-6 75 2483 664799 135852 135869 ACTGATTTTGAAGTCCCG 2815 2832 sooossssssssoooss 5-7-6 79 2048 664800 135853 135870 CACTGATTTTGAAGTCCC 2816 2833 sooossssssssoooss 5-7-6 82 2370 664534 135854 135871 TCACTGATTTTGAAGTCC 2817 2834 sooosssssssssooss 5-8-5 78 2372 664801 135854 135871 TCACTGATTTTGAAGTCC 2817 2834 sooossssssssoooss 5-7-6 29 2372 664802 135855 135872 ATCACTGATTTTGAAGTC 2818 2835 sooossssssssoooss 5-7-6 52 1668 664803 135856 135873 CATCACTGATTTTGAAGT 2819 2836 sooossssssssoooss 5-7-6 55 2377 664804 135857 135874 CCATCACTGATTTTGAAG 2820 2837 sooossssssssoooss 5-7-6 38 2380 664805 135858 135875 CCCATCACTGATTTTGAA 2821 2838 sooossssssssoooss 5-7-6 62 1669 664806 135859 135876 TCCCATCACTGATTTTGA 2822 2839 sooossssssssoooss 5-7-6 72 2385 664807 135860 135877 CTCCCATCACTGATTTTG 2823 2840 sooossssssssoooss 5-7-6 55 2388 664808 135861 135878 ACTCCCATCACTGATTTT 2824 2841 sooossssssssoooss 5-7-6 38 1670 664809 135862 135879 TACTCCCATCACTGATTT 2825 2842 sooossssssssoooss 5-7-6 54 2393 664810 135863 135880 TTACTCCCATCACTGATT 2826 2843 sooossssssssoooss 5-7-6 39 2396 664811 135864 135881 CTTACTCCCATCACTGAT 2827 2844 sooossssssssoooss 5-7-6 39 1671 664812 135865 135882 TCTTACTCCCATCACTGA 2828 2845 sooossssssssoooss 5-7-6 45 2401 664813 135866 135883 CTCTTACTCCCATCACTG 2829 2846 sooossssssssoooss 5-7-6 61 2404 664814 135867 135884 GCTCTTACTCCCATCACT 2830 2847 sooossssssssoooss 5-7-6 14 1672 664815 135868 135885 TGCTCTTACTCCCATCAC 2831 2848 sooossssssssoooss 5-7-6 70 2306 664816 135869 135886 TTGCTCTTACTCCCATCA 2832 2849 sooossssssssoooss 5-7-6 72 2307 664817 135870 135887 TTTGCTCTTACTCCCATC 2833 2850 sooossssssssoooss 5-7-6 62 1673 664818 135871 135888 ATTTGCTCTTACTCCCAT 2834 2851 sooossssssssoooss 5-7-6 69 2308 664819 135872 135889 AATTTGCTCTTACTCCCA 2835 2852 sooossssssssoooss 5-7-6 67 2309

TABLE 59 SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2 NO: 2 SEQ ISIS start stop start stop Linkage % ID NO site site Sequence site site Chemistry Motif inhibition NO 665045 21592 21609 ACATCCAAATAACAATAT n/a n/a sooosssssssssooss 5-8-5 1 2444 665046 21617 21634 TCCTGTCTCAAACACCTA n/a n/a sooosssssssssooss 5-8-5 44 2445 665047 30083 30100 AGCTTATCACAGTAGGTG n/a n/a sooosssssssssooss 5-8-5 94 2446 665048 30108 30125 CCTTTGCTTCACACACCA n/a n/a sooosssssssssooss 5-8-5 67 2447 621013 30133 30150 AGCTCACTACAGCAGGCA n/a n/a sooosssssssssooss 5-8-5 90 868 665049 30158 30175 TGGTTCTACAACCTGCCC n/a n/a sooosssssssssooss 5-8-5 57 2448 665050 30183 30200 CCTTTGTAACCTTGGAAC n/a n/a sooosssssssssooss 5-8-5 25 2449 665051 33838 33855 AGTTAAAAGATGGCAACC n/a n/a sooosssssssssooss 5-8-5 32 2450 665052 33863 33880 CTGGGCATTTTGATAAAA n/a n/a sooosssssssssooss 5-8-5 28 2451 621031 33888 33905 GGTATAATTTGTTTGGAC n/a n/a sooosssssssssooss 5-8-5 85 886 665053 33913 33930 AGTGCCAGACATTTTCAA n/a n/a sooosssssssssooss 5-8-5 20 2452 665054 33938 33955 CTGGCACAACAGGCATTG n/a n/a sooosssssssssooss 5-8-5 25 2453 621041 35721 35738 TTGCCATCTTGGACAGGG n/a n/a sooosssssssssooss 5-8-5 70 896 665055 35746 35763 GTGGTACAGAGGCTGTGT n/a n/a sooosssssssssooss 5-8-5 47 2454 665056 35771 35788 TTGCAGATTCTGAATACC n/a n/a sooosssssssssooss 5-8-5 78 2455 665057 64499 64516 TATGATTCCCAGGAGTCT n/a n/a sooosssssssssooss 5-8-5 0 2456 621181 64543 64560 GTAGGTGTCATCATCATC n/a n/a sooosssssssssooss 5-8-5 77 1113 665058 64568 64585 GTGACCCTTCAGGGCAAT n/a n/a sooosssssssssooss 5-8-5 35 2457 665059 64593 64610 ATGCAGGTGCCTTGCAGG n/a n/a sooosssssssssooss 5-8-5 36 2458 623965 73880 73897 TGATCTTCCATCACTTCG n/a n/a sooosssssssssooss 5-8-5 48 2278 665060 77364 77381 CCTGCTGCCAACAGTAGA n/a n/a sooosssssssssooss 5-8-5 51 2459 665061 77389 77406 CCCTCACGGGCTGTTGTG n/a n/a sooosssssssssooss 5-8-5 20 2460 621236 77414 77431 TCATCTGTGAAGCGGACG 73377 73394 sooosssssssssooss 5-8-5 79 706 665062 77439 77456 ACGGTCTGAGCATGAGGC n/a n/a sooosssssssssooss 5-8-5 77 2461 665063 77464 77481 AGCCATGGACCTGCTCTG n/a n/a sooosssssssssooss 5-8-5 48 2462 665064 77731 77748 GACAGTATACCCCACATC n/a n/a sooosssssssssooss 5-8-5 48 2463 621238 77781 77798 GTTATTTTGGAACAGTTT 73744 73761 sooosssssssssooss 5-8-5 89 708 665065 78142 78159 AAGAGTGGTAAAACCTAC n/a n/a sooosssssssssooss 5-8-5 51 2464 665066 80259 80276 GCTCACCAGCAGGAACTT n/a n/a sooosssssssssooss 5-8-5 56 2465 665067 80284 80301 AAGGGCCCAGTGTAGCAC n/a n/a sooosssssssssooss 5-8-5 66 2466 621251 80309 80326 AGTCATTATCATGTCACC 76272 76289 sooosssssssssooss 5-8-5 73 721 665068 80334 80351 CCAATAAGTGGCAGTGAT n/a n/a sooosssssssssooss 5-8-5 49 2467 665069 80359 80376 ACCATGCCTGGCAGATGA n/a n/a sooosssssssssooss 5-8-5 66 2468 665070 80799 80816 CTTAGTCATCTTCCCTCA n/a n/a sooosssssssssooss 5-8-5 64 2469 665071 80824 80841 ATTTCTGAATTCTTTGCC n/a n/a sooosssssssssooss 5-8-5 69 2470 621254 80849 80866 GGTAACATGTAAAGCTTC 76812 76829 sooosssssssssooss 5-8-5 76 724 665072 82508 82525 CTTACACTCTGGAAGGTT n/a n/a sooosssssssssooss 5-8-5 74 2471 665073 82533 82550 TATCCAGGGCTGAGCAGG n/a n/a sooosssssssssooss 5-8-5 38 2472 621263 82558 82575 ATGGGCTTATCAATGCAT 78521 78538 sooosssssssssooss 5-8-5 80 733 665074 82583 82600 GGAGTCAATCTGCCCTGG n/a n/a sooosssssssssooss 5-8-5 54 2473 665075 82608 82625 CTTCCCTGTGGCACTTTG n/a n/a sooosssssssssooss 5-8-5 29 2474 665076 89600 89617 ATGCCTCACTCAACAAGG n/a n/a sooosssssssssooss 5-8-5 26 2475 665077 89625 89642 CTCCACTTTGGGACCAGG n/a n/a sooosssssssssooss 5-8-5 87 2476 621302 89650 89667 GCTATGACCTAGTAGGAA n/a n/a sooosssssssssooss 5-8-5 68 772 665078 89675 89692 GGCACAGCACCCACATGC n/a n/a sooosssssssssooss 5-8-5 66 2477 665079 89700 89717 GGTGATAAAAGTTCACTA n/a n/a sooosssssssssooss 5-8-5 63 2531 620887 98891 98908 GTTTTCAAACACACCTTC n/a n/a sooosssssssssooss 5-8-5 91 665 98928 98945 664526 135842 135859 AAGTCCCGAGCCAAAGCC 2805 2822 sooosssssssssooss 5-8-5 55 2439 622116 135843 135860 GAAGTCCCGAGCCAAAGC 2806 2823 sooosssssssssooss 5-8-5 63 2045 664527 135844 135861 TGAAGTCCCGAGCCAAAG 2807 2824 sooosssssssssooss 5-8-5 40 2478 664528 135845 135862 TTGAAGTCCCGAGCCAAA 2808 2825 sooosssssssssooss 5-8-5 61 2479 622117 135846 135863 TTTGAAGTCCCGAGCCAA 2809 2826 sooosssssssssooss 5-8-5 55 2046 664529 135847 135864 TTTTGAAGTCCCGAGCCA 2810 2827 sooosssssssssooss 5-8-5 25 2480 664530 135848 135865 ATTTTGAAGTCCCGAGCC 2811 2828 sooosssssssssooss 5-8-5 38 2481 622118 135849 135866 GATTTTGAAGTCCCGAGC 2812 2829 sooosssssssssooss 5-8-5 63 2047 664531 135850 135867 TGATTTTGAAGTCCCGAG 2813 2830 sooosssssssssooss 5-8-5 66 2482 664532 135851 135868 CTGATTTTGAAGTCCCGA 2814 2831 sooosssssssssooss 5-8-5 73 2483 622119 135852 135869 ACTGATTTTGAAGTCCCG 2815 2832 sooosssssssssooss 5-8-5 66 2048 664533 135853 135870 CACTGATTTTGAAGTCCC 2816 2833 sooosssssssssooss 5-8-5 72 2370 664534 135854 135871 TCACTGATTTTGAAGTCC 2817 2834 sooosssssssssooss 5-8-5 61 2372 622120 135855 135872 ATCACTGATTTTGAAGTC 2818 2835 sooosssssssssooss 5-8-5 47 1668 664535 135856 135873 CATCACTGATTTTGAAGT 2819 2836 sooosssssssssooss 5-8-5 59 2377 664536 135857 135874 CCATCACTGATTTTGAAG 2820 2837 sooosssssssssooss 5-8-5 34 2380 622121 135858 135875 CCCATCACTGATTTTGAA 2821 2838 sooosssssssssooss 5-8-5 49 1669 664537 135859 135876 TCCCATCACTGATTTTGA 2822 2839 sooosssssssssooss 5-8-5 61 2385 664538 135860 135877 CTCCCATCACTGATTTTG 2823 2840 sooosssssssssooss 5-8-5 76 2388 622122 135861 135878 ACTCCCATCACTGATTTT 2824 2841 sooosssssssssooss 5-8-5 38 1670 664539 135862 135879 TACTCCCATCACTGATTT 2825 2842 sooosssssssssooss 5-8-5 57 2393 664540 135863 135880 TTACTCCCATCACTGATT 2826 2843 sooosssssssssooss 5-8-5 32 2396 622123 135864 135881 CTTACTCCCATCACTGAT 2827 2844 sooosssssssssooss 5-8-5 33 1671 664541 135865 135882 TCTTACTCCCATCACTGA 2828 2845 sooosssssssssooss 5-8-5 50 2401 664542 135866 135883 CTCTTACTCCCATCACTG 2829 2846 sooosssssssssooss 5-8-5 58 2404 622124 135867 135884 GCTCTTACTCCCATCACT 2830 2847 sooosssssssssooss 5-8-5 64 1672 623993 135868 135885 TGCTCTTACTCCCATCAC n/a n/a sooosssssssssooss 5-8-5 65 2306 623994 135869 135886 TTGCTCTTACTCCCATCA n/a n/a sooosssssssssooss 5-8-5 63 2307 622125 135870 135887 TTTGCTCTTACTCCCATC 2833 2850 sooosssssssssooss 5-8-5 75 1673 623995 135871 135888 ATTTGCTCTTACTCCCAT n/a n/a sooosssssssssooss 5-8-5 60 2308 623996 135872 135889 AATTTGCTCTTACTCCCA n/a n/a sooosssssssssooss 5-8-5 72 2309

TABLE 60 SEQ ID SEQ ID SEQ ID SEQ ID NO: 2 NO: SEQ ISIS NO: 1 NO: 1 start 2 stop % ID NO start site stop site Sequence site site Linkage Chemistry Motif inhibition NO 665043 73879 73897 TGATCTTCCATCACTTCGA 345 364 sooossssssssssooss 5-9-5 67 2484 665044 98890 98908 GTTTTCAAACACACCTTCA n/a n/a sooossssssssssooss 5-9-5 90 2485 98927 98945 665081 120039 120057 TTTTCTTACCACCCTAACA n/a n/a sooossssssssssooss 5-9-5 28 2486 665082 120041 120059 CGTTTTCTTACCACCCTAA n/a n/a sooossssssssssooss 5-9-5 86 2487 665083 120045 120063 AAACCGTTTTCTTACCACC n/a n/a sooossssssssssooss 5-9-5 86 2488 665084 120047 120065 AAAAACCGTTTTCTTACCA n/a n/a sooossssssssssooss 5-9-5 65 2489 665085 120060 120078 AGCTCATCAAAGCAAAAAC n/a n/a sooossssssssssooss 5-9-5 65 2490 665086 120110 120128 TCAAAAGACTATGTATTTT n/a n/a sooossssssssssooss 5-9-5 54 2491 665087 120389 120407 TGTAAATAATTGCCAAGTG n/a n/a sooossssssssssooss 5-9-5 61 2492 665088 120439 120457 TAAGCCACCATGCCTGTAA n/a n/a sooossssssssssooss 5-9-5 61 2493 665089 121711 121729 TGGACCCGCCTACTTGCTC n/a n/a sooossssssssssooss 5-9-5 85 2494 665090 121736 121754 TTTCGATGAGTGACATGCG n/a n/a sooossssssssssooss 5-9-5 50 2495 665091 121761 121779 TGCTTGCTCGCAAGGACGC n/a n/a sooossssssssssooss 5-9-5 80 2496 665092 121765 121783 CGCCTGCTTGCTCGCAAGG n/a n/a sooossssssssssooss 5-9-5 82 2497 665093 121767 121785 CCCGCCTGCTTGCTCGCAA n/a n/a sooossssssssssooss 5-9-5 92 2498 665094 121769 121787 GACCCGCCTGCTTGCTCGC n/a n/a sooossssssssssooss 5-9-5 91 2499 665095 121771 121789 TGGACCCGCCTGCTTGCTC n/a n/a sooossssssssssooss 5-9-5 83 2500 665096 121773 121791 CCTGGACCCGCCTGCTTGC n/a n/a sooossssssssssooss 5-9-5 88 2501 665097 121775 121793 ACCCTGGACCCGCCTGCTT n/a n/a sooossssssssssooss 5-9-5 60 2502 665098 121786 121804 AGTGACACGCCACCCTGGA n/a n/a sooossssssssssooss 5-9-5 62 2503 665099 121811 121829 CCTTTGGTAGCCAGAAAAA n/a n/a sooossssssssssooss 5-9-5 53 2504 665100 121817 121835 TCTGCACCTTTGGTAGCCA n/a n/a sooossssssssssooss 5-9-5 84 2505 665101 121938 121956 ACAGCACGGCGCATGGGAC n/a n/a sooossssssssssooss 5-9-5 54 2506 665102 121940 121958 CCACAGCACGGCGCATGGG n/a n/a sooossssssssssooss 5-9-5 84 2507 665103 121942 121960 AGCCACAGCACGGCGCATG n/a n/a sooossssssssssooss 5-9-5 85 2508 664714 135820 135838 TGACAAAAGCAGGTTAAGT 2783 2801 sooossssssssssooss 5-9-5 54 2332 664715 135821 135839 GTGACAAAAGCAGGTTAAG 2784 2802 sooossssssssssooss 5-9-5 65 2335 664716 135822 135840 AGTGACAAAAGCAGGTTAA 2785 2803 sooossssssssssooss 5-9-5 79 2337 664717 135823 135841 GAGTGACAAAAGCAGGTTA 2786 2804 sooossssssssssooss 5-9-5 70 2340 664718 135824 135842 CGAGTGACAAAAGCAGGTT 2787 2805 sooossssssssssooss 5-9-5 89 2343 664719 135825 135843 CCGAGTGACAAAAGCAGGT 2788 2806 sooossssssssssooss 5-9-5 84 2345 664720 135826 135844 GCCGAGTGACAAAAGCAGG 2789 2807 sooossssssssssooss 5-9-5 71 2348 664721 135827 135845 AGCCGAGTGACAAAAGCAG 2790 2808 sooossssssssssooss 5-9-5 81 2351 664722 135828 135846 AAGCCGAGTGACAAAAGCA 2791 2809 sooossssssssssooss 5-9-5 68 2353 664723 135829 135847 AAAGCCGAGTGACAAAAGC 2792 2810 sooossssssssssooss 5-9-5 51 2356 664724 135830 135848 CAAAGCCGAGTGACAAAAG 2793 2811 sooossssssssssooss 5-9-5 47 2359 664725 135831 135849 CCAAAGCCGAGTGACAAAA 2794 2812 sooossssssssssooss 5-9-5 69 2361 664726 135832 135850 GCCAAAGCCGAGTGACAAA 2795 2813 sooossssssssssooss 5-9-5 76 2364 664727 135833 135851 AGCCAAAGCCGAGTGACAA 2796 2814 sooossssssssssooss 5-9-5 64 2367 664728 135834 135852 GAGCCAAAGCCGAGTGACA 2797 2815 sooossssssssssooss 5-9-5 78 2369 664729 135835 135853 CGAGCCAAAGCCGAGTGAC 2798 2816 sooossssssssssooss 5-9-5 74 2421 664730 135836 135854 CCGAGCCAAAGCCGAGTGA 2799 2817 sooossssssssssooss 5-9-5 76 2424 664731 135837 135855 CCCGAGCCAAAGCCGAGTG 2800 2818 sooossssssssssooss 5-9-5 75 2426 664732 135838 135856 TCCCGAGCCAAAGCCGAGT 2801 2819 sooossssssssssooss 5-9-5 80 2429 664733 135839 135857 GTCCCGAGCCAAAGCCGAG 2802 2820 sooossssssssssooss 5-9-5 70 2432 664734 135840 135858 AGTCCCGAGCCAAAGCCGA 2803 2821 sooossssssssssooss 5-9-5 59 2434 664735 135841 135859 AAGTCCCGAGCCAAAGCCG 2804 2822 sooossssssssssooss 5-9-5 58 2437 664736 135842 135860 GAAGTCCCGAGCCAAAGCC 2805 2823 sooossssssssssooss 5-9-5 76 2440 664737 135843 135861 TGAAGTCCCGAGCCAAAGC 2806 2824 sooossssssssssooss 5-9-5 46 2442 664738 135844 135862 TTGAAGTCCCGAGCCAAAG 2807 2825 sooossssssssssooss 5-9-5 46 2550 664739 135845 135863 TTTGAAGTCCCGAGCCAAA 2808 2826 sooossssssssssooss 5-9-5 56 2551 664740 135846 135864 TTTTGAAGTCCCGAGCCAA 2809 2827 sooossssssssssooss 5-9-5 44 2552 664741 135847 135865 ATTTTGAAGTCCCGAGCCA 2810 2828 sooossssssssssooss 5-9-5 61 2553 664742 135848 135866 GATTTTGAAGTCCCGAGCC 2811 2829 sooossssssssssooss 5-9-5 59 2554 664743 135849 135867 TGATTTTGAAGTCCCGAGC 2812 2830 sooossssssssssooss 5-9-5 64 2555 664744 135850 135868 CTGATTTTGAAGTCCCGAG 2813 2831 sooossssssssssooss 5-9-5 67 2556 664745 135851 135869 ACTGATTTTGAAGTCCCGA 2814 2832 sooossssssssssooss 5-9-5 73 2557 664746 135852 135870 CACTGATTTTGAAGTCCCG 2815 2833 sooossssssssssooss 5-9-5 86 2558 664747 135853 135871 TCACTGATTTTGAAGTCCC 2816 2834 sooossssssssssooss 5-9-5 76 2371 664748 135854 135872 ATCACTGATTTTGAAGTCC 2817 2835 sooossssssssssooss 5-9-5 74 2373 664749 135855 135873 CATCACTGATTTTGAAGTC 2818 2836 sooossssssssssooss 5-9-5 50 2375 664750 135856 135874 CCATCACTGATTTTGAAGT 2819 2837 sooossssssssssooss 5-9-5 57 2378 664751 135857 135875 CCCATCACTGATTTTGAAG 2820 2838 sooossssssssssooss 5-9-5 64 2381 664752 135858 135876 TCCCATCACTGATTTTGAA 2821 2839 sooossssssssssooss 5-9-5 59 2383 664753 135859 135877 CTCCCATCACTGATTTTGA 2822 2840 sooossssssssssooss 5-9-5 73 2386 664754 135860 135878 ACTCCCATCACTGATTTTG 2823 2841 sooossssssssssooss 5-9-5 75 2389 664755 135861 135879 TACTCCCATCACTGATTTT 2824 2842 sooossssssssssooss 5-9-5 40 2391 664756 135862 135880 TTACTCCCATCACTGATTT 2825 2843 sooossssssssssooss 5-9-5 43 2394 664757 135863 135881 CTTACTCCCATCACTGATT 2826 2844 sooossssssssssooss 5-9-5 70 2397 664758 135864 135882 TCTTACTCCCATCACTGAT 2827 2845 sooossssssssssooss 5-9-5 61 2399 664759 135865 135883 CTCTTACTCCCATCACTGA 2828 2846 sooossssssssssooss 5-9-5 68 2402 664760 135866 135884 GCTCTTACTCCCATCACTG 2829 2847 sooossssssssssooss 5-9-5 82 2405 664761 135867 135885 TGCTCTTACTCCCATCACT 2830 2848 sooossssssssssooss 5-9-5 76 2407 664762 135868 135886 TTGCTCTTACTCCCATCAC 2831 2849 sooossssssssssooss 5-9-5 77 2409 664763 135869 135887 TTTGCTCTTACTCCCATCA 2832 2850 sooossssssssssooss 5-9-5 83 2411 664764 135870 135888 ATTTGCTCTTACTCCCATC 2833 2851 sooossssssssssooss 5-9-5 82 2413 664765 135871 135889 AATTTGCTCTTACTCCCAT 2834 2852 sooossssssssssooss 5-9-5 77 2415 664766 135872 135890 AAATTTGCTCTTACTCCCA 2835 2853 sooossssssssssooss 5-9-5 69 2417

TABLE 61 SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 NO: 1 NO: 2 NO: 2 ISIS start stop start stop NO site site Sequence site site Linkage Chemistry Motif % inhibition SEQ ID NO 665041 73879 73898 GTGATCTTCCATCACTTCGA  345  364 sooosssssssssssooss 5-10-5 86 25 665044 98890 98908 GTTTTCAAACACACCTTCA n/a n/a sooossssssssssooss 5-9-5 90 2485 98927 98945 665201 102119 102138 GGTTTGTTTTTTAAACAATT n/a n/a sooosssssssssssooss 5-10-5 57 2509 665202 102144 102163 GAACCCAATGAGAGTAGCAA n/a n/a sooosssssssssssooss 5-10-5 86 2510 665203 102169 102188 TTGCCAAAATCAGGAATGGG n/a n/a sooosssssssssssooss 5-10-5 87 2511 665204 102194 102213 TCAGGGCAATCTGGAAGCAT n/a n/a sooosssssssssssooss 5-10-5 79 2512 665205 102659 102678 ACTCTCCACTCCATGTCAAT n/a n/a sooosssssssssssooss 5-10-5 26 2513 665206 102684 102703 CAGAGCTCACAGCAATGATC n/a n/a sooosssssssssssooss 5-10-5 78 2514 665207 102709 102728 TCAGCATGAGTTGTGCCAAG n/a n/a sooosssssssssssooss 5-10-5 91 2515 665208 102734 102753 CCAGAGGAACTGTGTGCATT n/a n/a sooosssssssssssooss 5-10-5 79 2516 665209 102759 102778 CCAAGTTCCCTGAGGACATT n/a n/a sooosssssssssssooss 5-10-5 13 2517 665210 112934 112953 GTTGTGTTTTCTGGTTTATT n/a n/a sooosssssssssssooss 5-10-5 96 2518 665211 112959 112978 TTTTTTTTTAAGTTAGGAGT n/a n/a sooosssssssssssooss 5-10-5 20 2519 665212 112984 113003 TTTTACTGGTTGTGTTTTCT n/a n/a sooosssssssssssooss 5-10-5 90 2520 665213 113009 113028 CAGCTCTTAATGCTGTTATA n/a n/a sooosssssssssssooss 5-10-5 90 2521 665214 113498 113517 TTCAGCTCCTGCACCCAGCA n/a n/a sooosssssssssssooss 5-10-5 54 2522 665215 113548 113567 TTTTGCTTTTGCTCTGAAGA n/a n/a sooosssssssssssooss 5-10-5 57 2523 665216 115105 115124 TTGCTATTAAATATAATGTA n/a n/a sooosssssssssssooss 5-10-5 35 2524 665217 115130 115149 GCTTTTTAAAGTGACAACTG n/a n/a sooosssssssssssooss 5-10-5 78 2525 665218 115155 115174 TTCCACATAAATGTTCTACA n/a n/a sooosssssssssssooss 5-10-5 93 2526 665219 115205 115224 CCCTGATTGAGGAGAGGCAA n/a n/a sooosssssssssssooss 5-10-5 77 2527 665220 116595 116614 ATGTTTTTTTCTGGCCGGGC n/a n/a sooosssssssssssooss 5-10-5 85 2528 665221 116620 116639 TTCAGTATCTGCCACATACT n/a n/a sooosssssssssssooss 5-10-5 64 2529 665222 116645 116664 TGAATCAAAGGACATTAAGC n/a n/a sooosssssssssssooss 5-10-5 69 2530 664661 135820 135839 GTGACAAAAGCAGGTTAAGT 2783 2802 sooosssssssssssooss 5-10-5 78 2333 664662 135821 135840 AGTGACAAAAGCAGGTTAAG 2784 2803 sooosssssssssssooss 5-10-5 18 2336 664663 135822 135841 GAGTGACAAAAGCAGGTTAA 2785 2804 sooosssssssssssooss 5-10-5 79 2338 664664 135823 135842 CGAGTGACAAAAGCAGGTTA 2786 2805 sooosssssssssssooss 5-10-5 66 2341 664665 135824 135843 CCGAGTGACAAAAGCAGGTT 2787 2806 sooosssssssssssooss 5-10-5 94 2344 664666 135825 135844 GCCGAGTGACAAAAGCAGGT 2788 2807 sooosssssssssssooss 5-10-5 89 2346 664667 135826 135845 AGCCGAGTGACAAAAGCAGG 2789 2808 sooosssssssssssooss 5-10-5 29 2349 664668 135827 135846 AAGCCGAGTGACAAAAGCAG 2790 2809 sooosssssssssssooss 5-10-5 72 2352 664669 135828 135847 AAAGCCGAGTGACAAAAGCA 2791 2810 sooosssssssssssooss 5-10-5 53 2354 664670 135829 135848 CAAAGCCGAGTGACAAAAGC 2792 2811 sooosssssssssssooss 5-10-5 55 2357 664671 135830 135849 CCAAAGCCGAGTGACAAAAG 2793 2812 sooosssssssssssooss 5-10-5 58 2360 664672 135831 135850 GCCAAAGCCGAGTGACAAAA 2794 2813 sooosssssssssssooss 5-10-5 80 2362 664673 135832 135851 AGCCAAAGCCGAGTGACAAA 2795 2814 sooosssssssssssooss 5-10-5 52 2365 664674 135833 135852 GAGCCAAAGCCGAGTGACAA 2796 2815 sooosssssssssssooss 5-10-5 71 2368 664675 135834 135853 CGAGCCAAAGCCGAGTGACA 2797 2816 sooosssssssssssooss 5-10-5 76 2419 664676 135835 135854 CCGAGCCAAAGCCGAGTGAC 2798 2817 sooosssssssssssooss 5-10-5 78 2422 664677 135836 135855 CCCGAGCCAAAGCCGAGTGA 2799 2818 sooosssssssssssooss 5-10-5 77 2425 664678 135837 135856 TCCCGAGCCAAAGCCGAGTG 2800 2819 sooosssssssssssooss 5-10-5 73 2427 664679 135838 135857 GTCCCGAGCCAAAGCCGAGT 2801 2820 sooosssssssssssooss 5-10-5 73 2430 664680 135839 135858 AGTCCCGAGCCAAAGCCGAG 2802 2821 sooosssssssssssooss 5-10-5 20 2433 664681 135840 135859 AAGTCCCGAGCCAAAGCCGA 2803 2822 sooosssssssssssooss 5-10-5 64 2435 664682 135841 135860 GAAGTCCCGAGCCAAAGCCG 2804 2823 sooosssssssssssooss 5-10-5 60 2438 664683 135842 135861 TGAAGTCCCGAGCCAAAGCC 2805 2824 sooosssssssssssooss 5-10-5 64 2441 664684 135843 135862 TTGAAGTCCCGAGCCAAAGC 2806 2825 sooosssssssssssooss 5-10-5 52 2443 664685 135844 135863 TTTGAAGTCCCGAGCCAAAG 2807 2826 sooosssssssssssooss 5-10-5 45 2559 664686 135845 135864 TTTTGAAGTCCCGAGCCAAA 2808 2827 sooosssssssssssooss 5-10-5 56 2560 664687 135846 135865 ATTTTGAAGTCCCGAGCCAA 2809 2828 sooosssssssssssooss 5-10-5 67 2561 664688 135847 135866 GATTTTGAAGTCCCGAGCCA 2810 2829 sooosssssssssssooss 5-10-5 73 2562 664689 135848 135867 TGATTTTGAAGTCCCGAGCC 1644 1663 sooosssssssssssooss 5-10-5 67 56 664690 135849 135868 CTGATTTTGAAGTCCCGAGC 2812 2831 sooosssssssssssooss 5-10-5 73 464 664691 135850 135869 ACTGATTTTGAAGTCCCGAG 2813 2832 sooosssssssssssooss 5-10-5 85 2563 664692 135851 135870 CACTGATTTTGAAGTCCCGA 2814 2833 sooosssssssssssooss 5-10-5 89 2564 664693 135852 135871 TCACTGATTTTGAAGTCCCG 2815 2834 sooosssssssssssooss 5-10-5 75 2565 424880 135853 135872 ATCACTGATTTTGAAGTCCC 2816 2835 sssssssssssssssssss 5-10-5 62 57 664694 135853 135872 ATCACTGATTTTGAAGTCCC 1649 1668 sooosssssssssssooss 5-10-5 85 57 664695 135854 135873 CATCACTGATTTTGAAGTCC 2817 2836 sooosssssssssssooss 5-10-5 83 2374 664696 135855 135874 CCATCACTGATTTTGAAGTC 2818 2837 sooosssssssssssooss 5-10-5 66 2376 664697 135856 135875 CCCATCACTGATTTTGAAGT 2819 2838 sooosssssssssssooss 5-10-5 58 2379 664698 135857 135876 TCCCATCACTGATTTTGAAG 2820 2839 sooosssssssssssooss 5-10-5 71 2382 664699 135858 135877 CTCCCATCACTGATTTTGAA 2821 2840 sooosssssssssssooss 5-10-5 32 2384 664700 135859 135878 ACTCCCATCACTGATTTTGA 2822 2841 sooosssssssssssooss 5-10-5 71 2387 664701 135860 135879 TACTCCCATCACTGATTTTG 2823 2842 sooosssssssssssooss 5-10-5 75 2390 664702 135861 135880 TTACTCCCATCACTGATTTT 2824 2843 sooosssssssssssooss 5-10-5 46 2392 664703 135862 135881 CTTACTCCCATCACTGATTT 2825 2844 sooosssssssssssooss 5-10-5 22 2395 664704 135863 135882 TCTTACTCCCATCACTGATT 2826 2845 sooosssssssssssooss 5-10-5 72 2398 664705 135864 135883 CTCTTACTCCCATCACTGAT 2827 2846 sooosssssssssssooss 5-10-5 70 2400 664706 135865 135884 GCTCTTACTCCCATCACTGA 2828 2847 sooosssssssssssooss 5-10-5 77 2403 664707 135866 135885 TGCTCTTACTCCCATCACTG 2829 2848 sooosssssssssssooss 5-10-5 88 2406 664708 135867 135886 TTGCTCTTACTCCCATCACT 2830 2849 sooosssssssssssooss 5-10-5 86 2408 664709 135868 135887 TTTGCTCTTACTCCCATCAC 2831 2850 sooosssssssssssooss 5-10-5 16 2410 664710 135869 135888 ATTTGCTCTTACTCCCATCA 2832 2851 sooosssssssssssooss 5-10-5 80 2412 664711 135870 135889 AATTTGCTCTTACTCCCATC 2833 2852 sooosssssssssssooss 5-10-5 65 2414 664712 135871 135890 AAATTTGCTCTTACTCCCAT 2834 2853 sooosssssssssssooss 5-10-5 60 2416 664713 135872 135891 GAAATTTGCTCTTACTCCCA 2835 2854 sooosssssssssssooss 5-10-5 85 2418

Example 17 Dose-Dependent Antisense Inhibition of Human Tau in SH-SY5Y Cells

Gapmers from studies described above exhibiting significant in vitro inhibition of tau mRNA were selected and tested at various doses in SH-SY5Y cells. The antisense oligonucleotides were tested in a series of experiments that had similar culture conditions. The results for each experiment are presented in separate tables shown below. Cells were plated at a density of 20,000 cells per well and transfected using electroporation with 0.247 μM, 0.741 μM, 2.22 μM, 6.67 μM, and 20.00 μM concentrations of antisense oligonucleotide, as specified in the Tables below. After a treatment period of approximately 16 hours, RNA was isolated from the cells and tau mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS3104 was used to measure mRNA levels. Tau mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of tau, relative to untreated control cells. Tau mRNA levels were significantly reduced in a dose-dependent manner in antisense oligonucleotide treated cells.

TABLE 62 0.247 0.741 2.22 6.67 20.00 IC₅₀ ISIS No μM μM μM μM μM (μM) 620887 18 39 71 88 95 1.2 664662 5 16 34 59 68 6.4 664665 16 41 69 89 96 1.2 664667 5 20 39 78 90 2.9 664680 7 11 21 56 81 5.9 664699 0 1 28 59 85 5.7 664709 8 16 48 66 89 3.1 665044 15 33 59 84 94 1.6 665205 0 7 35 51 86 5.3 665207 22 36 57 82 93 1.4 665210 15 47 74 94 97 1.0 665212 6 22 56 82 95 2.1 665213 0 18 57 78 93 2.5 665217 10 28 63 86 95 1.7 665218 10 31 64 85 93 1.7

TABLE 63 0.247 0.741 2.22 6.67 20.00 IC₅₀ ISIS No μM μM μM μM μM (μM) 664718 26 34 60 88 86 1.3 664745 34 39 67 87 95 0.9 664746 15 37 60 87 92 1.5 664760 32 50 75 89 97 0.7 665044 21 44 75 92 97 0.9 665082 5 27 67 87 96 1.7 665083 17 34 67 85 97 1.4 665089 9 26 53 84 94 2.0 665093 11 57 81 93 95 0.9 665094 25 53 76 91 93 0.7 665095 20 33 63 89 95 1.4 665096 5 37 57 86 95 1.8 665102 8 36 61 90 96 1.6 665103 6 24 59 83 92 2.1

TABLE 64 0.247 0.741 2.22 6.67 20.00 IC₅₀ ISIS No μM μM μM μM μM (μM) 620887 26 45 72 89 96 1.2 621013 26 49 82 93 97 0.7 621031 25 41 66 84 92 1.1 621041 26 55 70 89 92 0.7 621238 23 41 59 85 95 1.2 621251 33 56 74 88 92 0.5 664514 11 36 68 79 91 1.6 664516 16 36 64 82 94 1.5 664534 18 35 48 76 91 1.9 664775 24 30 59 79 92 1.6 664788 11 21 26 55 73 6.2 664799 26 37 63 79 90 1.3 664800 15 0 61 75 90 2.7 665047 43 65 82 95 98 0.2 665077 19 44 63 85 96 1.2

Example 18 Intracerebroventricular Administration of Antisense Oligonucleotides Against Human Tau mRNA in Htau Mice

Selected compounds were tested for efficacy by ICV administration in human tau transgenic mice (Duff et al., Neurobiology of Disease 7:87-98, 2000).

Treatment and Surgery

Groups of 4 mice each were administered ISIS 613099, ISIS 613361, ISIS 613370, ISIS 623782, or ISIS 623996 with a 200 μg dose delivered by ICV bolus injection. A control group of 2 mice was similarly treated with ISIS 424880 and a control group of 4 mice was similarly treated with PBS. All procedures were performed under isoflourane anesthesia and in accordance with IACUC regulations. For mouse ICV bolus injections, the antisense oligonucleotide was injected into the right lateral ventricle of human tau transgenic mice. Ten microliters of solution containing 200 μg of oligonucleotide in PBS was injected over approximately 10 seconds. Tissue was collected 14 days after oligonucleotide administration.

RNA Analysis

On day 14 after the oligonucleotide administration, RNA was extracted from the hippocampus, spinal cord and cortex for real-time PCR analysis of tau mRNA levels. Human tau mRNA levels were measured using the human primer probe set RTS3104. Results were calculated as percent inhibition of human tau mRNA expression compared to the control. All the antisense oligonucleotides effect significant inhibition of human tau mRNA levels in several tissues.

TABLE 65 Percent reduction of human tau mRNA levels in hTau mice ISIS Spinal No Cortex Hippocampus Cord 613099 41.8 55.2 54.1 613361 69.4 57.3 72.5 613370 42.2 61.0 63.0 623782 62.2 79.1 70.8 623996 74.1 69.2 76.4 424880 50.3 53.1 55.2

Example 19 Design of Oligonucleotides Targeting Human Tau

ISIS No. 603054 was designed to target human Tau. The nucleobase sequence and linkage chemistry of ISIS No. 603054 is given in table 66 below. ISIS No. 603054 is a 5-10-5 MOE gapmer. ISIS No. 603054 is 20 nucleosides in length, wherein the central gap segment comprises ten 2′-deoxynucleosides and is flanked by wing segments on the 5′ direction and the 3′ direction comprising five nucleosides each. Each nucleoside in the 5′ wing segment and each nucleoside in the 3′ wing segment has a 2′-MOE modification. All cytosine residues throughout each gapmer are 5-methylcytosines. “Start site” indicates the 5 ‘-most nucleoside to which the gapmer is targeted in the human gene sequence. “Stop site” indicates the 3’-most nucleoside to which the gapmer is targeted human gene sequence. Each gapmer listed in Table 1 below is targeted to either the human tau genomic sequence, designated herein as SEQ ID NO: 1 (GENBANK Accession No. NT_010783.15 truncated from nucleotides 9240000 to 9381000) or to the human tau mRNA sequence, designated herein as SEQ ID NO: 2 (GENBANK Accession No. NM_001123066.3).

TABLE 66 ICV in vivo study in hTau mice and WT C57Bl6 mice SEQ SEQ ID ID SEQ ID SEQ ID NO: 2 NO: 2 SEQ ISIS NO: 1 NO: 1 start stop ID NO start site stop site Sequence site site Linkage Chemistry NO 603054 135853 135872 ATCACTGATTTTGAAGTCCC 2816 2835 sooosssssssssssooos 57

Example 20 In Vivo Analysis in Mice of Oligonucleotides Targeting Human Tau

Oligonucleotides, shown in the table below, were designed to target Tau. Mice, either human tau transgenic mice “hTau” (Duff et al., Neurobiology of Disease 7:87-98, 2000; Davies et al. J. Neurochem. (2003) 86, 582-590) or wild-type WT C57B16 mice were separated into groups of 3 or 4 mice. Each mouse in each group of mice was administered a single ICV dose of either 300 ug or 200 ug each of the oligonucleotides in the table below. At 3 hours post injection, each mouse was evaluated according to 7 different criteria. The 7 criteria are (1) the mouse was bright, alert, and responsive; (2) the mouse was standing or hunched without stimuli; (3) the mouse shows any movement without stimuli (4) the mouse demonstrates forward movement after its lifted; (5) the mouse demonstrates any movement after its lifted; (6) the mouse responds to a tail pinch; (7) regular breathing. For each of the 7 different criteria, each mouse was given a sub-score of 0 if it met the criteria or 1 if it did not. After all of the 7 criteria were evaluated, the sub-scores were summed for each mouse and then averaged for each group. For example, if a mouse was bright, alert, and responsive 3 hours after the 300 μg ICV dose, and met all other criteria, it would get a summed score of 0. If another mouse was not bright, alert, and responsive 3 hours after the 300 μg ICV dose but met all other criteria, it would receive a score of 1. Saline treated mice generally receive a score of 0. Results are presented as the average score for each treatment group in Table 67 below. “ND” means no data. These results demonstrate that ISIS 613099, ISIS 613361, ISIS 613370, ISIS 623782, ISIS 623996, ISIS 424880, and ISIS 603054 were well tolerated.

TABLE 67 ICV in vivo study in hTau mice and WT C57Bl6 mice Line: hTau WT C57Bl6 Dose: 300 ug 200 ug 300 ug 613099 0 ND ND 613361 0 ND ND 613370 0 ND ND 623782 0 ND ND 623996 0 ND ND 424880 ND 3 ND 603054 ND ND 0.25

Example 20 In Vivo Analysis in Rats of Oligonucleotides Targeting Human Tau

Sprague Dawley rats were separated into groups of 4 rats each. Each rat in each group of rats was administered a single 1 mg intrathecal (IT) dose or a single 3 mg intrathecal (IT) dose of ISIS 613099, ISIS 613361, ISIS 613370, ISIS 623782, ISIS 623996, ISIS 424880, or ISIS 603054. At 3 hours after injection, the movement of 7 different parts of the body was evaluated for each rat. The 7 body parts are (1) the rat's tail; (2) the rat's posterior posture; (3) the rat's hind limbs; (4) the rat's hind paws; (5) the rat's forepaws; (6) the rat's anterior posture; and (7) the rat's head. For each of the 7 different body parts, each rat was given a sub-score of 0 if the body part was moving or 1 if the body part was paralyzed. After each of the 7 body parts were evaluated, the sub-scores were summed for each rat and then averaged for each group. For example, if a rat's tail, head, and all other evaluated body parts were moving 3 hours after the 3 mg IT dose, it would get a summed score of 0. If another rat was not moving its tail 3 hours after the 3 mg IT dose but all other evaluated body parts were moving, it would receive a score of 1. Saline treated rats generally receive a score of 0. A score of at the top end of the range would be suggestive of toxicity. Results are presented as the average score for each treatment group in Table 68 below.

TABLE 68 1 mg and 3 mg IT bolus in vivo study Score 3 hours after injection ISIS Dose No. 1 mg 3 mg 613099 0.25 3 613361 0 0.33 613370 0 0 623782 0.25 0.75 623996 0 2 424880 2.3 6 603054 1.25 2.75 

What is claimed is:
 1. A compound comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides and having a nucleobase sequence comprising at least 8 contiguous nucleobases of a sequence selected from among SEQ ID Nos:1699, 1700, 1701, 1702, and 1703, wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO:
 1. 2. The compound of claim 1, wherein at least one internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage.
 3. The compound of claim 2, wherein the modified oligonucleotide is a single-stranded oligonucleotide.
 4. The compound of claim 2, wherein the modified internucleoside linkage is a phosphorothioate internucleoside linkage.
 5. The compound of claim 4, wherein at least one internucleoside linkage of the modified oligonucleotide is a phosphodiester internucleoside linkage.
 6. The compound of claim 2, wherein at least one nucleobase of the modified oligonucleotide is a modified nucleobase.
 7. The compound of claim 6, wherein the modified nucleobase is a 5-methylcytosine.
 8. The compound of claim 1, wherein at least one nucleoside of the modified oligonucleotide comprises a modified sugar.
 9. The compound of claim 8, wherein at least one modified sugar is a bicyclic sugar.
 10. The compound of claim 9, wherein each bicyclic sugar comprises a chemical bridge between the 4′ and 2′ positions of the sugar, wherein each chemical bridge is independently selected from: 4′-CH(R)—O-2′ and 4′-(CH₂)₂—O-2′, wherein each R is independently selected from H, C₁-C₆ alkyl, and C₁-C₆ alkoxy.
 11. The compound of claim 10, wherein at least one chemical bridge is 4′-CH(R)—O-2′ and wherein R is methyl.
 12. The compound of claim 10, wherein at least one chemical bridge is 4′-CH(R)—O-2′ and wherein R is H.
 13. The compound of claim 10, wherein at least one chemical bridge is 4′-CH(R)—O-2′ and wherein R is —CH₂—O—CH₃.
 14. The compound of claim 7, wherein at least one modified sugar comprises a 2′-O-methoxyethyl group or a 2′-O-methyl group.
 15. The compound of claim 1, wherein the at least one nucleoside of the modified oligonucleotide comprises a sugar surrogate.
 16. The compound of claim 15, wherein the at least one sugar surrogate is a morpholino or a peptide nucleic acid.
 17. The compound of claim 1, wherein the modified oligonucleotide comprises: a 5′ wing segment having from 1 to 6 nucleosides and wherein each nucleoside of the 5′ wing segment comprises a modified sugar; a 3′ wing segment having from 1 to 6 nucleosides and wherein each nucleoside of the 3′ wing segment comprises a modified sugar; and a gap segment having from 8 to 15 nucleosides and wherein each nucleoside of the gap segment is a deoxynucleoside.
 18. The compound of claim 1, wherein the compound comprises a conjugate group.
 19. A composition comprising a compound according to claim 1 and a pharmaceutically acceptable carrier or diluent.
 20. A compound comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides and comprising a nucleobase sequence having at least 8 consecutive nucleobases complementary to an equal length portion of nucleobases 139811 to 139836 of SEQ ID NO: 1, wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO:
 1. 21. The compound of claim 20, wherein the nucleobase sequence has at least 12 consecutive nucleobases selected from among SEQ ID Nos: 1699 -1703.
 22. The compound of claim 20, wherein at least one internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage.
 23. The compound of claim 22, wherein the modified oligonucleotide is a single-stranded oligonucleotide.
 24. The compound of claim 22, wherein the modified internucleoside linkage is a phosphorothioate internucleoside linkage.
 25. The compound of claim 24, wherein at least one internucleoside linkage of the modified oligonucleotide is a phosphodiester internucleoside linkage.
 26. The compound of claim 22, wherein at least one nucleobase of the modified oligonucleotide is a modified nucleobase.
 27. The compound of claim 26, wherein the modified nucleobase is a 5-methylcytosine.
 28. The compound of claim 20, wherein at least one nucleoside of the modified oligonucleotide comprises a modified sugar.
 29. The compound of claim 28, wherein at least one modified sugar is a bicyclic sugar.
 30. The compound of claim 29, wherein each bicyclic sugar comprises a chemical bridge between the 4′ and 2′ positions of the sugar, wherein each chemical bridge is independently selected from: 4′-CH(R)—O-2′ and 4′-(CH₂)₂—O-2′, wherein each R is independently selected from H, C₁-C₆ alkyl, and C₁-C₆ alkoxy.
 31. The compound of claim 30, wherein at least one chemical bridge is 4′-CH(R)—O-2′ and wherein R is methyl.
 32. The compound of claim 30, wherein at least one chemical bridge is 4′-CH(R)—O-2′ and wherein R is H.
 33. The compound of claim 30, wherein at least one chemical bridge is 4′-CH(R)—O-2′ and wherein R is —CH₂—O—CH₃.
 34. The compound of claim 28, wherein at least one modified sugar comprises a 2′-O-methoxyethyl group or a 2′-O-methyl group.
 35. The compound of claim 20, wherein the at least one nucleoside of the modified oligonucleotide comprises a sugar surrogate.
 36. The compound of claim 35, wherein the at least one sugar surrogate is a morpholino or a peptide nucleic acid.
 37. The compound of claim 20, wherein the modified oligonucleotide comprises: a 5′ wing segment having from 1 to 6 nucleosides and wherein each nucleoside of the 5′ wing segment comprises a modified sugar; a 3′ wing segment having from 1 to 6 nucleosides and wherein each nucleoside of the 3′ wing segment comprises a modified sugar; and a gap segment having from 8 to 15 nucleosides and wherein each nucleoside of the gap segment is a deoxynucleoside.
 38. The compound of claim 20, wherein the compound comprises a conjugate group.
 39. A composition comprising a compound according to claim 20 and a pharmaceutically acceptable carrier or diluent.
 40. The compound of claim 1, wherein the modified oligonucleotide comprises: a 5′ wing segment having from 1 to 6 nucleosides and wherein at least 4 nucleosides of the 5′ wing segment comprises a modified sugar; a 3′ wing segment having from 1 to 6 nucleosides and wherein at least 4 nucleosides of the 3′ wing segment comprises a modified sugar; and a gap segment having from 8 to 15 nucleosides and wherein each nucleoside of the gap segment is a deoxynucleoside.
 41. The compound of claim 40, wherein 1 or 2 nucleosides of the 5′ wing segment is a deoxynucleoside and 1 or 2 nucleosides of the 3′ wing segment is a deoxynucleoside.
 42. The compound of claim 41, wherein at least one modified sugar is a bicyclic sugar.
 43. The compound of claim 42, wherein each bicyclic sugar comprises a chemical bridge between the 4′ and 2′ positions of the sugar, wherein each chemical bridge is independently selected from: 4′-CH(R)—O-2′ and 4′-(CH₂)₂—O-2′, wherein each R is independently selected from H, C₁-C₆ alkyl, and C₁-C₆ alkoxy.
 44. The compound of claim 20, wherein the modified oligonucleotide comprises: a 5′ wing segment having from 1 to 6 nucleosides and wherein at least 4 nucleosides of the 5′ wing segment comprises a modified sugar; a 3′ wing segment having from 1 to 6 nucleosides and wherein at least 4 nucleosides of the 3′ wing segment comprises a modified sugar; and a gap segment having from 8 to 15 nucleosides and wherein each nucleoside of the gap segment is a deoxynucleoside.
 45. The compound of claim 44, wherein 1 or 2 nucleosides of the 5′ wing segment is a deoxynucleoside and 1 or 2 nucleosides of the 3′ wing segment is a deoxynucleoside.
 46. The compound of claim 45, wherein at least one modified sugar is a bicyclic sugar.
 47. The compound of claim 46, wherein each bicyclic sugar comprises a chemical bridge between the 4′ and 2′ positions of the sugar, wherein each chemical bridge is independently selected from: 4′-CH(R)—O-2′ and 4′-(CH₂)₂—O-2′, wherein each R is independently selected from H, C₁-C₆ alkyl, and C₁-C₆ alkoxy. 